Details of the Researcher

PHOTO

Shinichi Fukushige
Section
Graduate School of Medicine
Job title
Specially Appointed Professor(Research)
Degree

  • 医学博士（大阪大学）

Education 1

  • Osaka University　Graduate School, Division of Medicine　生理系

    - 1988/03

Committee Memberships 4

  • International Journal of Molecular Sciences　Editorial Board Member

    2020/10 - Present

  • Scientifica　Academic Editor

    2012/01 - Present

  • Epigenetic Diagnosis & Therapy　編集委員

    2014/03 - 2018/12

  • Genetics Research International　編集委員

    2010/06 - 2015/12

Professional Memberships 3

  • American Association for Cancer Research(2000/04-)

  • 日本分子生物学会(1985/08-)

  • 日本癌学会(1996/03-)

Research Interests 2

  • DNA mismatch repair

  • Cancer Epigenetics

Research Areas 1

  • Life sciences / Experimental pathology / Molecular Biology

Awards 2

  1. 平成２３年度東北大学医学部奨学賞金賞

    2012/01/13　東北大学医学部　癌におけるDNA高度メチル化を指標とするバイオマーカーの探索

  2. 宮城県医師会医学奨励賞

    2012/01/04　宮城県医師会

Papers 96

  1. Association between NLRP3 Inflammasome and Tumor-Node-Metastasis Staging in Prostate Cancer: Immunohistochemical Studies of Prostate Needle Biopsy and Radical Prostatectomy Specimens. Peer-reviewed

    Toshiya Miyauchi, Shintaro Narita, Yuriko Saiki, Yukitsugu Kudo-Asabe, Akira Horii, Shinichi Fukushige, Tomonori Habuchi, Hiroshi Nanjo, Akiteru Goto

    The Tohoku journal of experimental medicine　2024/08/01

    DOI： 10.1620/tjem.2024.J074 　

  2. TWIST1 is a prognostic factor for neoadjuvant chemotherapy for patients with resectable pancreatic cancer: a preliminary study. Peer-reviewed

    Sho Fujiwara, Yuriko Saiki, Shinichi Fukushige, Mie Yamanaka, Masaharu Ishida, Fuyuhiko Motoi, Michiaki Unno, Akira Horii

    Surgery today　2023/02/10

    DOI： 10.1007/s00595-023-02655-3 　

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    Recent advances in the development of chemotherapies have helped improve the prognosis of pancreatic ductal adenocarcinoma (PDAC). However, predicting factors for the outcomes of chemotherapies (either gemcitabine or S-1) have not yet been established. We analyzed the expression of 4 major epithelial-to-mesenchymal transition-inducing transcription factors in 38 PDAC patients who received adjuvant chemotherapy after radical resection to examine the association with patients' prognoses. The TWIST1-positive group showed a significantly poorer prognosis than the TWIST1-negative group for both the relapse-free survival (median survival time [MST] of 8.9 vs. 18.5 months, P = 0.016) and the overall survival (MST of 15.2 vs. 33.4 months, P = 0.023). A multivariate analysis revealed that TWIST1 positivity was an independent prognostic factor for a poor response to adjuvant chemotherapies (hazard ratio 2.61; 95% confidence interval 1.10-6.79; P = 0.029). These results suggest that TWIST1 can be utilized as an important poor prognostic factor for radically resected PDAC patients with adjuvant chemotherapy, potentially including neoadjuvant therapy using these agents.

  3. Long Non-Coding RNAs Associated with Mitogen-Activated Protein Kinase in Human Pancreatic Cancer Peer-reviewed

    Tomohiko Ishikawa, Shinichi Fukushige, Yuriko Saiki, Katsuya Hirose, Takako Hiyoshi, Takenori Ogawa, Yukio Katori, Toru Furukawa

    Cancers　15　(1)　303-303　2023/01/02

    Publisher: MDPI AG

    DOI： 10.3390/cancers15010303 　

    eISSN： 2072-6694

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    Long non-coding RNAs (lncRNAs) have emerged as a significant player in various cancers, including pancreatic cancer. However, how lncRNAs are aberrantly expressed in cancers is largely unknown. We hypothesized that lncRNAs would be regulated by signaling pathways and contribute to malignant phenotypes of cancer. In this study, to understand the significance of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK), which is a major aberrant signaling pathway in pancreatic cancer, for the expression of lncRNAs, we performed comparative transcriptome analyses between pancreatic cancer cell lines with or without activation of MAPK. We identified 45 lncRNAs presumably associated with MAPK in pancreatic cancer cells; among these, LINC00941 was consistently upregulated by MAPK. The immediate genomic upstream region flanking LINC00941 was identified as a promoter region, the activity of which was found to be preferentially associated with MAPK activity via ETS-1 binding site. LINC00941 promoted cell proliferation in vitro. Moreover, TCGA data analysis indicated that high expression of LINC00941 was associated with poor prognosis of patients with pancreatic cancer. Transcriptomes comparing transcriptions between cells with and without LINC00941 knockdown revealed 3229 differentially expressed genes involved in 44 biological processes, including the glycoprotein biosynthetic process, beta-catenin-TCF complex assembly, and histone modification. These results indicate that MAPK mediates the aberrant expression of lncRNAs. LINC00941 is the lncRNA by MAPK most consistently promoted, and is implicated in the dismal prognosis of pancreatic cancer. MAPK-associated lncRNAs may play pivotal roles in malignant phenotypes of pancreatic cancer, and as such might represent both potentially valid therapeutic targets and diagnostic biomarkers.

  4. Combination therapy of lymphatic drug delivery and total-body irradiation in a metastatic lymph node and lung mouse model. International-journal Peer-reviewed

    Shota Sora, Ariunbuyan Sukhbaatar, Shinichi Fukushige, Shiro Mori, Maya Sakamoto, Tetsuya Kodama

    Cancer science　2022/09/03

    DOI： 10.1111/cas.15562 　

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    Chemotherapy using a lymphatic drug delivery system (LDDS) targeting lymph nodes (LNs) in the early stage of metastasis has a superior antitumor effect to systemic chemotherapy. LDDS produces a higher drug retention rate and tissue selectivity in LNs. To expand the therapeutic coverage of LDDS from local treatment of metastatic LNs to prevention of distant metastases, the combination of treatment with therapies that enhance systemic tumor immune effects is an important therapeutic strategy. Recently, low-dose total-body irradiation (TBI) has been shown to activate immune responses and alter the tumor microenvironment. Here we show that combination therapy with TBI and LDDS improves the antitumor effect of metastatic LNs and lung metastasis. Tumor cells were inoculated into the subiliac LN to induce metastasis into the mouse proper axillary LN (PALN) and lung. Total-body irradiation was conducted on day 4 after inoculation using a gamma irradiator. LDDS was administrated into the accessory axillary LN to treat PALN. In vivo bioluminescence imaging system, high-frequency ultrasound system, and histology showed that combination therapy using TBI (total dose 1.0 Gy once) and the LDDS suppressed tumor growth in LNs and lung metastases and was more effective than using LDDS or TBI alone. Quantitative RT-PCR of spleens after combination therapy revealed increased expression of CD4, CD8, and IL-12b, indicating an activated immune response. The results show that combination therapy with TBI and LDDS is a method to improve the efficacy of LN metastases and distant metastases therapy and is a promising novel approach to treat cancer patients.

  5. Development of a system combining comprehensive genotyping and organoid cultures for identifying and testing genotype-oriented personalised medicine for pancreatobiliary cancers International-journal Peer-reviewed

    Masahiro Shiihara, Tomohiko Ishikawa, Yuriko Saiki, Yuko Omori, Katsuya Hirose, Shinichi Fukushige, Naoki Ikari, Ryota Higuchi, Masakazu Yamamoto, Takanori Morikawa, Kei Nakagawa, Hiroki Hayashi, Masamichi Mizuma, Hideo Ohtsuka, Fuyuhiko Motoi, Michiaki Unno, Yasunobu Okamura, Kengo Kinoshita, Toru Furukawa

    European Journal of Cancer　148　239-250　2021/05

    Publisher: Elsevier BV

    DOI： 10.1016/j.ejca.2021.01.047 　

    ISSN： 0959-8049

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    BACKGROUND: Pancreatobiliary cancer is a highly aggressive tumour with a dismal prognosis. Personalised medicine represents a promising and effective therapeutic approach for this intractable disease. In this study, we aimed to establish a system for identifying and testing genotype-oriented targeted drugs for pancreatobiliary cancers by combining exome sequencing and organoid culture of primary tumours. METHODS: Tumour cells isolated from resected tumours were subjected to organoid cultures based on published protocols with modifications. Exome sequencing was performed on the primary tumours. Histopathological and molecular features of the primary tumours were validated in the corresponding organoids. Genotype-oriented candidate targeted drugs were identified from exome sequencing, and their efficacies were tested in the organoids. RESULTS: Organoid cultures succeeded in 30 of 54 (55.6%) cases. Six primary cancers of the biliary tract and gall bladder were subjected to exome sequencing, which revealed a variety of somatic mutations of genes involved in signalling pathways, epigenetic modifiers, genome maintenance and metabolic enzymes. Most of the organoids of these 6 cases showed identical histopathological features and genomic aberrations as those of the primary tumours. Some of the aberrations were candidates for targeted therapies. Integrin-linked kinase (ILK) was one such candidate target, and an ILK inhibitor was confirmed to suppress proliferation of patient-derived organoids. CONCLUSIONS: By combining exome sequencing and organoid culture, our model enabled to identify genotype-oriented targets for personalised medicine and to test efficacies of candidate targeted drugs in the organoids. The current proof-of-concept approach could increase therapeutic opportunities for patients with pancreatobiliary cancers.

  6. Aberrant Hypermethylation-Mediated Suppression of PYCARD Is Extremely Frequent in Prostate Cancer with Gleason Score ≥ 7. International-journal Peer-reviewed

    Toshiya Miyauchi, Masahiro Takahashi, Koji Mitsuzuka, Yuriko Saiki, Teppei Okubo, Paula M Vertino, Akiteru Goto, Yoichi Arai, Akira Horii, Shinichi Fukushige

    Disease Markers　2021　8858905-8858905　2021

    DOI： 10.1155/2021/8858905 　

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    Epigenetic gene silencing by aberrant DNA methylation leads to loss of key cellular pathways in tumorigenesis. In order to analyze the effects of DNA methylation on prostate cancer, we established LNCaP-derived human prostate cancer cells that can pharmacologically induce global reactivation of hypermethylated genes by the methyl-CpG targeted transcriptional activation (MeTA) method. The MeTA suppressed the growth of LNCaP-derived cells and induced apoptosis. Microarray analysis indicated that PYCARD (PYD and CARD domain containing) encoding an apoptosis-inducing factor was upregulated by 65-fold or more after treatment with MeTA. We analyzed DNA methylation statuses using 50 microdissected primary prostate cancer tissues and found an extremely high frequency of tumor-specific promoter hypermethylation of PYCARD (90%, 45/50). Moreover, DNA methylation status was significantly associated with Gleason score (P = 0.0063); the frequency of tumor-specific hypermethylation was 96% (44/46) in tumors with Gleason score ≥ 7, whereas that in tumors with Gleason score 6 was 25% (1/4). Immunohistochemical analyses using these 50 cases indicated that only 8% (4/50) of cancerous tissues expressed PYCARD, whereas 80% (40/50) of corresponding normal prostate epithelial and/or basal cells expressed PYCARD. In addition, there was no relationship between PYCARD immunostaining and the Gleason score in cancerous tissue and surrounding normal tissue. Inducible expression of PYCARD inhibited cell proliferation by induction of apoptosis. These results suggest that aberrant methylation of PYCARD is a distinctive feature of prostate cancers with Gleason score ≥ 7 and may play an important role in escaping from apoptosis in prostatic tumorigenesis.

  7. Epigenetic inactivation of IRX4 is responsible for acceleration of cell growth in human pancreatic cancer. International-journal Peer-reviewed

    Kanchan Chakma, Zhaodi Gu, Yakefujiang Abudurexiti, Tatsuo Hata, Fuyuhiko Motoi, Michiaki Unno, Akira Horii, Shinichi Fukushige

    Cancer Science　111　(12)　4594-4604　2020/09/07

    DOI： 10.1111/cas.14644 　

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    Epigenetic gene silencing by aberrant DNA methylation is one of the important mechanisms leading to loss of key cellular pathways in tumorigenesis. Methyl-CpG targeted transcriptional activation (MeTA) reactivates hypermethylation-mediated silenced genes in a different way from DNA demethylating agents. Microarray coupled with MeTA (MeTA-array) identified seven commonly hypermethylation-mediated silenced genes in 12 pancreatic ductal adenocarcinoma (PDAC) cell lines. Among these, we focused on IRX4 (Iroquois homeobox 4) because IRX4 is located at chromosome 5p15.33 where one of PDAC susceptibility loci has been identified through genome-wide association study. IRX4 was greatly downregulated in all of the analyzed 12 PDAC cell lines by promoter hypermethylation. In addition, the IRX4 promoter region was found to be frequently and specifically hypermethylated in primary resected PDACs (18/28: 64%). Re-expression of IRX4 inhibited colony formation and proliferation in two PDAC cell lines, PK-1 and PK-9. In contrast, knockdown of IRX4 accelerated cell proliferation in an IRX4-expressing normal pancreatic ductal epithelial cell line, HPDE-1. Because IRX4 is a sequence-specific transcription factor, downstream molecules of IRX4 were pursued by microarray analyses utilizing tetracycline-mediated IRX4 inducible PK-1 and PK-9 cells; CRYAB, CD69, and IL32 were identified as IRX4-downstream candidate genes. Forced expression of these genes suppressed colony formation abilities for both PK-1 and PK-9. These results suggest that DNA methylation-mediated silencing of IRX4 contributes to pancreatic tumorigenesis through aberrant transcriptional regulation of several cancer-related genes.

  8. Methylation-mediated silencing of the LIM homeobox 6 (LHX6) gene promotes cell proliferation in human pancreatic cancer. International-journal Peer-reviewed

    Yakefujiang Abudurexiti, Zhaodi Gu, Kanchan Chakma, Tatsuo Hata, Fuyuhiko Motoi, Michiaki Unno, Akira Horii, Shinichi Fukushige

    Biochemical and Biophysical Research Communications　526　(3)　626-632　2020/04/02

    DOI： 10.1016/j.bbrc.2020.03.120 　

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    Epigenetic gene silencing by aberrant DNA methylation leads to loss of key cellular pathways in tumorigenesis. DNA methylation-mediated silenced genes in pancreatic cancer were searched for using the methyl-CpG targeted transcriptional activation (MeTA) method, and LHX6 (LIM homeobox 6), a transcription factor involved in embryogenesis and head development, was selected as a strong candidate gene. LHX6 was downregulated in most of the pancreatic cancer cell lines (83%, 10/12), mainly through promoter hypermethylation and histone deacetylation. Furthermore, LHX6 was methylated in primary pancreatic cancer specimens (57%, 16/28) in a tumor-specific manner. Re-expression of LHX6 inhibited colony formation and proliferation in LHX6 low-expressing pancreatic cancer cell lines, PK-1 and PK-9. In contrast, knockdown of LHX6 accelerated cell proliferation in LHX6 high-expressing pancreatic cancer cell lines, PCI-35 and MIA PaCa-2. In order to analyze LHX6 downstream genes, we performed microarray analyses using LHX6 inducible PK-1 and PK-9 and found that LHX6 induction upregulated several genes that had tumor suppressive functions. Among these, we focused on TFPI2 (Tissue factor pathway inhibitor-2) and found that TFPI2 was greatly downregulated in all twelve pancreatic cancer cell lines. Our present results suggest that epigenetic inactivation of LHX6 plays an important role in pancreatic tumorigenesis by promoting cell proliferation through aberrant transcriptional regulation of several cancer-related genes.

  9. CD45+CD326+ Cells are Predictive of Poor Prognosis in Non-Small Cell Lung Cancer Patients. International-journal Peer-reviewed

    Kota Ishizawa, Mie Yamanaka, Yuriko Saiki, Eisaku Miyauchi, Shinichi Fukushige, Tetsuya Akaishi, Atsuko Asao, Takahiro Mimori, Ryota Saito, Yutaka Tojo, Riu Yamashita, Michiaki Abe, Akira Sakurada, Nhu-An Pham, Ming Li, Yoshinori Okada, Tadashi Ishii, Naoto Ishii, Seiichi Kobayashi, Masao Nagasaki, Masakazu Ichinose, Ming-Sound Tsao, Akira Horii

    Clinical cancer research : an official journal of the American Association for Cancer Research　25　(22)　6756-6763　2019/11/15

    DOI： 10.1158/1078-0432.CCR-19-0545 　

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    PURPOSE: The epithelial-to-mesenchymal transition, the major process by which some cancer cells convert from an epithelial phenotype to a mesenchymal one, has been suggested to drive chemo-resistance and/or metastasis in patients with cancer. However, only a few studies have demonstrated the presence of CD45/CD326 doubly-positive cells (CD45/CD326 DPC) in cancer. We deployed a combination of cell surface markers to elucidate the phenotypic heterogeneity in non-small cell lung cancer (NSCLC) cells and identified a new subpopulation that is doubly-positive for epithelial and non-epithelial cell-surface markers in both NSCLC cells and patients' malignant pleural effusions. EXPERIMENTAL DESIGN: We procured a total of 39 patients' samples, solid fresh lung cancer tissues from 21 patients and malignant pleural effusion samples from 18 others, and used FACS and fluorescence microscopy to check their surface markers. We also examined the EGFR mutations in patients with known acquired EGFR mutations. RESULTS: Our data revealed that 0.4% to 17.9% of the solid tumor tissue cells and a higher percentage of malignant pleural effusion cells harbored CD45/CD326 DPC expressing both epithelial and nonepithelial surface markers. We selected 3 EGFR mutation patients and genetically confirmed that the newly identified cell population really originated from cancer cells. We also found that higher proportions of CD45/CD326 DPC are significantly associated with poor prognosis. CONCLUSIONS: In conclusion, varying percentages of CD45/CD326 DPC exist in both solid cancer tissue and malignant pleural effusion in patients with NSCLC. This CD45/CD326 doubly-positive subpopulation can be an important key to clinical management of patients with NSCLC.

  10. Expression of SNAIL in accompanying PanIN is a key prognostic indicator in pancreatic ductal adenocarcinomas. International-journal Peer-reviewed

    Fujiwara S, Saiki Y, Ishizawa K, Fukushige S, Yamanaka M, Sato M, Ishida M, Motoi F, Unno M, Horii A.

    Cancer Med　8　(4)　1671-1678　2019

    DOI： 10.1002/cam4.2016 　

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    Pancreatic ductal adenocarcinoma (PDAC) is the most lethal cancer, mainly because of its invasive and metastatic characteristics. Pancreatic intraepithelial neoplasia (PanIN) is one of the major precursor lesions of PDAC. Although epithelial-to-mesenchymal transition (EMT) is known to play an important role for these malignant behaviors, the association between PanIN and EMT has not been clearly understood. Therefore, we explored possible molecules for regulation of EMT immunohistochemically. Using surgically resected specimens from 71 PDAC patients, expressions of SNAIL, SLUG, TWIST1, and ZEB1 were investigated in high-grade PanIN (HG-PanIN) and PDAC. Results demonstrated that PDAC accompanied by SNAIL-positive HG-PanIN showed a significantly better relapse-free survival (RFS) (median survival time (MST) of 11.3 months vs 4.4 months, P < 0.001) and overall survival overall survival (OS) (MST of 25.2 months vs 13.6 months, P < 0.001). In PDAC accompanied by SLUG-positive HG-PanIN, RFS and OS (P = 0.09 and P = 0.05) tended to have a better prognosis. In contrast, we could not find any significant prognostic benefits in the expression of TWIST1 or ZEB1 in PDAC accompanied by HG-PanIN. Our present results suggest that (1) EMT may play an important role in the development of PDAC from HG-PanIN, and (2) SNAIL may predict a distinct subgroup that shows a better prognosis.

  11. S100A10 upregulation associates with poor prognosis in lung squamous cell carcinoma. Peer-reviewed

    Sato K, Saiki Y, Arai K, Ishizawa K, Fukushige S, Aoki K, Abe J, Takahashi S, Sato I, Sakurada A, Okada Y, Horii A.

    Biochem Biophys Res Commun　505　466-470　2018/09

  12. NDRG2, suppressed expression associates with poor prognosis in pancreatic cancer, is hypermethylated in the second promoter in human gastrointestinal cancers Peer-reviewed

    Akihiro Yamamura, Koh Miura, Hideaki Karasawa, Fuyuhiko Motoi, Yasuhiko Mizuguchi, Yuriko Saiki, Shinichi Fukushige, Makoto Sunamura, Chikashi Shibata, Michiaki Unno, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　484　(1)　138-143　2017/02

    DOI： 10.1016/j.bbrc.2017.01.055 　

    ISSN： 0006-291X

    eISSN： 1090-2104

  13. Atrial natriuretic peptide induces peroxisome proliferator activated receptor γ during cardiac ischemia-reperfusion in swine heart. Peer-reviewed

    Tomoyuki Suzuki, Yuriko Saiki, Akira Horii, Shinichi Fukushige, Shunsuke Kawamoto, Osamu Adachi, Masatoshi Akiyama, Koki Ito, Naoki Masaki, Yoshikatsu Saiki

    General thoracic and cardiovascular surgery　65　(2)　85-95　2017/02

    DOI： 10.1007/s11748-016-0704-6 　

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    OBJECTIVES: Atrial natriuretic peptide is a cardiac atrium-derived hormone and its cardioprotective effects have recently been confirmed, but the actual mechanism underlying these effects has not been well elucidated. In this study, we proposed that atrial natriuretic peptide achieves its effects in part via peroxisome proliferator activated receptor γ, a nuclear receptor. METHODS: Hemodynamic data in swine heart ischemia-reperfusion model were measured under the conditions of no medication for control (Group N, n = 8) or that of carperitide (synthetic human atrial natriuretic peptide) systemic administration (Group A, n = 8). After 30 min of left anterior descending artery total occlusion and 4 h of reperfusion, peroxisome proliferator activated receptor γ mRNA and protein expressions in cardiac muscle were examined. The mRNA expression of Liver X receptor α, the downstream agent of peroxisome proliferator activated receptor γ, was also evaluated. Creatine kinase-myocardial band and Troponin T elevations after reperfusion were evaluated as markers of cardiac damage. RESULTS: The dP/dT decrease during reperfusion was ameliorated in Group A. Peroxisome proliferator activated receptor γ mRNA expression in Group A was significantly higher in ischemic area than that in Group N, although the difference was not significant in the marginal and non-ischemic areas. The peroxisome proliferator activated receptor γ protein expression in ischemic area was also significantly dominant in Group A. CONCLUSIONS: Atrial natriuretic peptide may achieve its cardioprotective effects in part via the activation of the peroxisome proliferator activated receptor γ pathway, particularly in central areas of ischemic lesions.

  14. Targeted TET oxidase activity through methyl-CpG-binding domain extensively suppresses cancer cell proliferation Peer-reviewed

    Yasuhiko Mizuguchi, Yuriko Saiki, Akira Horii, Shinichi Fukushige

    CANCER MEDICINE　5　(9)　2522-2533　2016/09

    DOI： 10.1002/cam4.830 　

    ISSN： 2045-7634

  15. Single-dose rosuvastatin ameliorates lung ischemia-reperfusion injury via upregulation of endothelial nitric oxide synthase and inhibition of macrophage infiltration in rats with pulmonary hypertension Peer-reviewed

    Satoshi Matsuo, Yuriko Saiki, Osamu Adachi, Shunsuke Kawamoto, Shinichi Fukushige, Akira Horii, Yoshikatsu Saiki

    JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY　149　(3)　902-909　2015/03

    DOI： 10.1016/j.jtcvs.2014.10.030 　

    ISSN： 0022-5223

    eISSN： 1097-685X

  16. DNA methylation as a biomarker in cancer Invited Peer-reviewed

    Shinichi Fukushige, Akira Horii

    Biomarkers in Disease: Methods, Discoveries and Applications: Biomarkers in Cancer　107-133　2015/01/01

    Publisher: Springer Netherlands

    DOI： 10.1007/978-94-007-7681-4_45 　

  17. Technological advances in epigenomics lead to a better understanding of inflammatory diseases, decitabine and H3K27me3. International-journal Invited Peer-reviewed

    Shinichi Fukushige, Akira Horii

    Epigenomics　7　(2)　133-6　2015

    Publisher: Future Medicine Ltd.

    DOI： 10.2217/epi.14.90 　

    ISSN： 1750-192X 1750-1911

  18. S100A4 is frequently overexpressed in lung cancer cells and promotes cell growth and cell motility Peer-reviewed

    Na Chen, Daisuke Sato, Yuriko Saiki, Makoto Sunamura, Shinichi Fukushige, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　447　(3)　459-464　2014/05

    DOI： 10.1016/j.bbrc.2014.04.025 　

    ISSN： 0006-291X

    eISSN： 1090-2104

  19. Road to early detection of pancreatic cancer: Attempts to utilize epigenetic biomarkers Invited Peer-reviewed

    Shinichi Fukushige, Akira Horii

    CANCER LETTERS　342　(2)　231-237　2014/01

    DOI： 10.1016/j.canlet.2012.03.022 　

    ISSN： 0304-3835

    eISSN： 1872-7980

  20. Suppressed expression of NDRG2 correlates with poor prognosis in pancreatic cancer Peer-reviewed

    Akihiro Yamamura, Koh Miura, Hideaki Karasawa, Kazuhiro Morishita, Keiko Abe, Yasuhiko Mizuguchi, Yuriko Saiki, Shinichi Fukushige, Naoyuki Kaneko, Tomohiko Sase, Hiroki Nagase, Makoto Sunamura, Fuyuhiko Motoi, Shinichi Egawa, Chikashi Shibata, Michiaki Unno, Iwao Sasaki, Akira Horii

    Biochemical and Biophysical Research Communications　441　(1)　102-107　2013/11/08

    DOI： 10.1016/j.bbrc.2013.10.010 　

    ISSN： 0006-291X 1090-2104

  21. DNA methylation in cancer: a gene silencing mechanism and the clinical potential of its biomarkers. Invited Peer-reviewed

    Shinichi Fukushige, Akira Horii

    The Tohoku Journal of Experimental Medicine　229　(3)　173-85　2013/03

    DOI： 10.1620/tjem.229.173 　

    ISSN： 0040-8727

    eISSN： 1349-3329

  22. S100A4, frequently overexpressed in various human cancers, accelerates cell motility in pancreatic cancer cells Peer-reviewed

    Hitoshi Sekine, Na Chen, Keisuke Sato, Yuriko Saiki, Yuki Yoshino, Yukiko Umetsu, Guo Jin, Hiroki Nagase, Zhaodi Gu, Shinichi Fukushige, Makoto Sunamura, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　429　(3-4)　214-219　2012/12

    DOI： 10.1016/j.bbrc.2012.10.048 　

    ISSN： 0006-291X

  23. miR-34a is downregulated in cis-diamminedichloroplatinum treated sinonasal squamous cell carcinoma patients with poor prognosis Peer-reviewed

    Takenori Ogawa, Yuriko Saiki, Kiyoto Shiga, Na Chen, Shinichi Fukushige, Makoto Sunamura, Hiroki Nagase, Sho Hashimoto, Kazuto Matsuura, Shigeru Saijo, Toshimitsu Kobayashi, Akira Horii

    CANCER SCIENCE　103　(9)　1737-1743　2012/09

    DOI： 10.1111/j.1349-7006.2012.02338.x 　

    ISSN： 1347-9032

  24. DCK is frequently inactivated in acquired gemcitabine-resistant human cancer cells Peer-reviewed

    Yuriko Saiki, Yuki Yoshino, Hiroko Fujimura, Tatsuya Manabe, Yuki Kudo, Miki Shimada, Nariyasu Mano, Tomohiro Nakano, Yoonha Lee, Shinjiro Shimizu, Shinya Oba, Sho Fujiwara, Hideyuki Shimizu, Na Chen, Zhaleh Kashkouli Nezhad, Guo Jin, Shinichi Fukushige, Makoto Sunamura, Masaharu Ishida, Fuyuhiko Motoi, Shinichi Egawa, Michiaki Unno, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　421　(1)　98-104　2012/04

    DOI： 10.1016/j.bbrc.2012.03.122 　

    ISSN： 0006-291X

  25. Identification of epigenetically silenced genes in human pancreatic cancer by a novel method "microarray coupled with methyl-CpG targeted transcriptional activation" (MeTA-array) Peer-reviewed

    Hideyuki Shimizu, Akira Horii, Makoto Sunamura, Fuyuhiko Motoi, Shinichi Egawa, Michiaki Unno, Shinichi Fukushige

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　411　(1)　162-167　2011/07

    DOI： 10.1016/j.bbrc.2011.06.121 　

    ISSN： 0006-291X

  26. Microarray coupled with methyl-CpG targeted transcriptional activation (MeTA-array) identifies hypermethylated genes containing the stringent criteria of CpG islands at high frequency Peer-reviewed

    Yuko Sato, Akira Horii, Shinichi Fukushige

    EPIGENETICS　6　(6)　752-759　2011/06

    DOI： 10.4161/epi.6.6.15906 　

    ISSN： 1559-2294

  27. Methyl-CpG targeted recruitment of p300 reactivates tumor suppressor genes in human cancer cells Peer-reviewed

    Shinichi Fukushige, Emiko Kondo, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　379　(4)　1021-1026　2009/02

    DOI： 10.1016/j.bbrc.2009.01.010 　

    ISSN： 0006-291X

  28. Methyl-CpG targeted transcriptional activation allows re-expression of tumor suppressor genes in human cancer cells Peer-reviewed

    Shinichi Fukushige, Emiko Kondo, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　377　(2)　600-605　2008/12

    DOI： 10.1016/j.bbrc.2008.10.016 　

    ISSN： 0006-291X

  29. Involvement of CREM in CYP1A1 induction through ligand-independent activation pathway of aryl hydrocarbon receptor in HepG2 cells Peer-reviewed

    Eriko Sasamori, Shuji Shimoyama, Shinichi Fukushige, Hideaki Kikuchi

    ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS　478　(1)　26-35　2008/10

    DOI： 10.1016/j.abb.2008.07.021 　

    ISSN： 0003-9861

  30. The interplay between hMLH1 and hMRE11: Role in MMR and the effect of hMLH1 mutations Peer-reviewed

    Nianxi Zhao, Fengxue Zhu, Fenghua Yuan, Anoria K. Haick, Shinichi Fukushige, Liya Gu, Chengtao Her

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　370　(2)　338-343　2008/05

    DOI： 10.1016/j.bbrc.2008.03.082 　

    ISSN： 0006-291X

  31. RET finger protein enhances MBD2-and MBD4-dependent transcriptional repression Peer-reviewed

    Shinichi Fukushige, Emiko Kondo, Zhaodi Gu, Hideyuki Suzuki, Akira Horii

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS　351　(1)　85-92　2006/12

    DOI： 10.1016/j.bbrc.2006.10.005 　

    ISSN： 0006-291X

  32. The thymine DNA glycosylase MBD4 represses transcription and is associated with methylated p16(INK4a) and hMLH1 genes Peer-reviewed

    E Kondo, ZD Gu, A Horii, S Fukushige

    MOLECULAR AND CELLULAR BIOLOGY　25　(11)　4388-4396　2005/06

    DOI： 10.1128/MCB.25.11.4388-4396.2005 　

    ISSN： 0270-7306

  33. Comparative genomic hybridization and mutation analyses of sporadic schwannomas Peer-reviewed

    T Ikeda, S Hashimoto, S Fukushige, H Ohmori, A Horii

    JOURNAL OF NEURO-ONCOLOGY　72　(3)　225-230　2005/05

    DOI： 10.1007/s11060-004-2693-z 　

    ISSN： 0167-594X

  34. The ability of Sgs1 to interact with DNA topoisomerase III is essential for damage-induced recombination Peer-reviewed

    A Ui, M Seki, H Ogiwara, R Onodera, S Fukushige, F Onoda, T Enomoto

    DNA REPAIR　4　(2)　191-201　2005/02

    DOI： 10.1016/j.dnareps.2004.09.002 　

    ISSN： 1568-7864

  35. The role of chromosome 18 abnormalities in the progression of pancreatic adenocarcinoma Peer-reviewed

    M Sunamura, LP Lefter, DG Duda, R Morita, H Inoue, T Yokoyama, T Yatsuoka, T Abe, S Egawa, T Furukawa, S Fukushige, M Oshimura, A Horii, S Matsuno

    PANCREAS　28　(3)　311-316　2004/04

    DOI： 10.1097/00006676-200404000-00019 　

    ISSN： 0885-3177

  36. DSCP1, a novel TP53-inducible gene, is upregulated by strong genotoxic stresses and its overexpression inhibits tumor cell growth in vitro Peer-reviewed

    T Hata, T Ogawa, TA Yokoyama, S Fukushige, A Horii, T Furukawa

    INTERNATIONAL JOURNAL OF ONCOLOGY　24　(3)　513-520　2004/03

    ISSN： 1019-6439

  37. A yeast two-hybrid assay provides a simple way to evaluate the vast majority of hMLH1 germ-line mutations Peer-reviewed

    E Kondo, H Suzuki, A Horii, S Fukushige

    CANCER RESEARCH　63　(12)　3302-3308　2003/06

    ISSN： 0008-5472

  38. Presence on human chromosome 10 of omeprazole-sensitivity gene whose product mediates CYP1A1 induction Peer-reviewed

    H Kikuchi, S Fukushige, M Shibazaki, Y Shiratori

    CYTOGENETIC AND GENOME RESEARCH　97　(1-2)　51-57　2002

    DOI： 10.1159/000064040 　

    ISSN： 1424-8581

  39. The interacting domains of three MutL heterodimers in man: hMLH1 interacts with 36 homologous amino acid residues within hMLH3, hPMS1 and hPMS2 Peer-reviewed

    E Kondo, A Horii, S Fukushige

    NUCLEIC ACIDS RESEARCH　29　(8)　1695-1702　2001/04

    DOI： 10.1093/nar/29.8.1695 　

    ISSN： 0305-1048

  40. Isolation and characterization of the human gene homologous to the Drosophila headcase (hdc) gene in chromosome bands 6q23-q24, a region of common deletion in human pancreatic cancer Peer-reviewed

    N Makino, T Yamato, H Inoue, T Furukawa, T Abe, T Yokoyama, T Yatsuoka, S Fukushige, S Orikasa, T Takahashi, A Horii

    DNA SEQUENCE　11　(6)　547-553　2001

    DOI： 10.3109/10425170109041340 　

    ISSN： 1042-5179

  41. Identification of three commonly deleted regions on chromosome arm 6q in human pancreatic cancer. (vol 25, pg 60, 1999)

    T Abe, N Makino, T Furukawa, H Ouyang, M Kimura, T Yatsuoka, T Yokoyama, H Inoue, S Fukushige, M Hoshi, Y Hayashi, M Sunamura, M Kobari, S Matsuno, A Horii

    GENES CHROMOSOMES & CANCER　29　(1)　88-88　2000/09

    ISSN： 1045-2257

  42. Not hMSH2 but hMLH1 is frequently silenced by hypermethylation in endometrial cancer but rarely silenced in pancreatic cancer with microsatellite instability Peer-reviewed

    E Kondo, T Furukawa, K Yoshinaga, H Kijima, S Semba, T Yatsuoka, T Yokoyama, S Fukushige, A Horii

    INTERNATIONAL JOURNAL OF ONCOLOGY　17　(3)　535-541　2000/09

    ISSN： 1019-6439

  43. Frequent loss of copy number on the long arm of chromosome 21 in human esophageal squamous cell carcinoma Peer-reviewed

    T Mayama, S Fukushige, R Shineha, T Nishihira, S Satomi, A Horii

    INTERNATIONAL JOURNAL OF ONCOLOGY　17　(2)　245-252　2000/08

    ISSN： 1019-6439

  44. Association of poor prognosis with loss of 12q, 17p, and 18q, and concordant loss of 6q/17p and 12q/18q in human pancreatic ductal adenocarcinoma Peer-reviewed

    T Yatsuoka, M Sunamura, T Furukawa, S Fukushige, T Yokoyama, H Inoue, K Shibuya, K Takeda, S Matsuno, A Horii

    AMERICAN JOURNAL OF GASTROENTEROLOGY　95　(8)　2080-2085　2000/08

    DOI： 10.1016/S0002-9270(00)00963-1 　

    ISSN： 0002-9270

  45. p24/ING1-ALT1 and p47/ING1-ALT2, distinct alternative transcripts of p33/ING1 Peer-reviewed

    A Saito, T Furukawa, S Fukushige, S Koyama, M Hoshi, Y Hayashi, A Horii

    JOURNAL OF HUMAN GENETICS　45　(3)　177-181　2000

    DOI： 10.1007/s100380050206 　

    ISSN： 1434-5161

  46. Adenovirus-mediated delivery of the PTEN gene inhibits cell growth by induction of apoptosis in endometrial cancer Peer-reviewed

    A Sakurada, H Hamada, S Fukushige, T Yokoyama, K Yoshinaga, T Furukawa, S Sato, A Yajima, M Sato, S Fujimura, A Horii

    INTERNATIONAL JOURNAL OF ONCOLOGY　15　(6)　1069-1074　1999/12

    ISSN： 1019-6439

  47. 膵癌で高頻度の欠失の見られた第6番染色体長腕の癌抑制遺伝子の解析

    井上 寛子, 牧野 直彦, 阿部 忠義, 古川 徹, 八岡 利昌, 横山 忠明, 福重 真一, 砂村 真琴, 武田 和憲, 堀井 明

    膵臓　14　(4)　340-340　1999/09

    Publisher: (一社)日本膵臓学会

    ISSN： 0913-0071

    eISSN： 1881-2805

  48. Identification of a 5-cM region of common allelic loss on 8p12-p21 in human breast cancer and genomic analysis of the hEXT1L/EXTR1/EXTL3 gene in this locus Peer-reviewed

    A Suzuki, XY Shao, XQ Song, T Hanaoka, S Irie, M Kashiwada, G Samara, LG Close, T Aoki, M Fujimori, Y Ishikawa, M Hatori, M Hosaka, A Sakurada, M Sato, N Ohuchi, S Satomi, S Fukushige, A Horii, T Sato

    INTERNATIONAL JOURNAL OF ONCOLOGY　15　(3)　443-451　1999/09

    ISSN： 1019-6439

  49. 膵がんで高頻度に欠失している第12番染色体長腕の解析

    古川 徹, 八岡 利昌, Youssef Emile M, 福重 真一, 砂村 真琴, 堀井 明

    日本癌学会総会記事　58回　127-127　1999/08

    Publisher: (一社)日本癌学会

    ISSN： 0546-0476

  50. Allelic imbalanceを指標にした膵癌の存在診断及び悪性度診断法の検討

    八岡 利昌, 古川 徹, 福重 真一, 木村 光宏, 阿部 忠義, 砂村 眞琴, 武田 和憲, 松野 正紀, 堀井 明

    日本癌学会総会記事　58回　127-127　1999/08

    Publisher: (一社)日本癌学会

    ISSN： 0546-0476

  51. Identification of three commonly deleted regions on chromosome arm 6q in human pancreatic cancer Peer-reviewed

    T Abe, N Makino, T Furukawa, H Ouyang, M Kimura, T Yatsuoka, T Yokoyama, H Inoue, S Fukushige, M Hoshi, Y Hayashi, M Sunamura, M Kobari, S Matsuno, A Horii

    GENES CHROMOSOMES & CANCER　25　(1)　60-64　1999/05

    DOI： 10.1002/(SICI)1098-2264(199905)25:1<60::AID-GCC9>3.0.CO;2-Y 　

    ISSN： 1045-2257

  52. Chromosome band 16q24 is frequently deleted in human gastric cancer Peer-reviewed

    Y Mori, M Matsunaga, T Abe, S Fukushige, K Miura, M Sunamura, K Shiiba, M Sato, T Nukiwa, A Horii

    BRITISH JOURNAL OF CANCER　80　(3-4)　556-562　1999/05

    DOI： 10.1038/sj.bjc.6690391 　

    ISSN： 0007-0920

  53. The human PMS2L proteins do not interact with hMLH1, a major DNA mismatch repair protein Peer-reviewed

    E Kondo, A Horii, S Fukushige

    JOURNAL OF BIOCHEMISTRY　125　(4)　818-825　1999/04

    ISSN： 0021-924X

  54. Infrequent somatic mutations of the p73 gene in various human cancers Peer-reviewed

    S Han, S Semba, T Abe, N Makino, T Furukawa, S Fukushige, H Takahashi, A Sakurada, M Sato, K Shiibai, S Matsuno, Y Nimura, A Nakagawara, A Horii

    EUROPEAN JOURNAL OF SURGICAL ONCOLOGY　25　(2)　194-198　1999/04

    DOI： 10.1053/ejso.1998.0626 　

    ISSN： 0748-7983

  55. Cloning and characterization of the human UDP-N-acetylglucosamine: alpha-1,3-D-mannoside beta-1,4-N-acetylglucosaminyltransferase IV-homologue (hGnT-IV-H) gene Peer-reviewed

    T Furukawa, EM Youssef, T Yatsuoka, T Yokoyama, N Makino, H Inoue, S Fukushige, M Hoshi, Y Hayashi, M Sunamura, A Horii

    JOURNAL OF HUMAN GENETICS　44　(6)　397-401　1999

    DOI： 10.1007/s100380050186 　

    ISSN： 1434-5161

  56. Isolation and characterization of the novel gene, TU3A, in a commonly deleted region on 3p14.3 -&gt; p14.2 in renal cell carcinoma Peer-reviewed

    T Yamato, K Orikasa, S Fukushige, S Orikasa, A Horii

    CYTOGENETICS AND CELL GENETICS　87　(3-4)　291-295　1999

    ISSN： 0301-0171

  57. Loss of chromosome 18q is an early event in pancreatic ductal tumorigenesis Peer-reviewed

    S Fukushige, T Furukawa, K Satoh, M Sunamura, M Kobari, M Koizumi, A Horii

    CANCER RESEARCH　58　(19)　4222-4226　1998/10

    ISSN： 0008-5472

  58. ヒト膵癌における6q21の500-kbの共通欠失領域の詳細な解析

    阿部 忠義, 牧野 直彦, 古川 徹, 福重 真一, 八岡 利昌, 砂村 真琴, 小針 雅男, 松野 正紀, 堀井 明

    日本癌学会総会記事　57回　133-133　1998/08

    Publisher: (一社)日本癌学会

    ISSN： 0546-0476

  59. 膵臓癌における12q21上の共通欠失領域の解析

    エミール・M・ヨセフ, 古川 徹, 八岡 利昌, 福重 真一, 砂村 真琴, 小針 雅男, 松野 正紀, 堀井 明

    日本癌学会総会記事　57回　133-133　1998/08

    Publisher: (一社)日本癌学会

    ISSN： 0546-0476

  60. 膵癌の発生・進展に関与する癌抑制遺伝子 12q22-q23.1の650-kbの共通欠失領域からの遺伝子同定

    八岡 利昌, 古川 徹, 阿部 忠義, 木村 光宏, 福重 真一, 砂村 眞琴, 小針 雅男, 松野 正紀, 堀井 明

    日本癌学会総会記事　57回　470-470　1998/08

    Publisher: (一社)日本癌学会

    ISSN： 0546-0476

  61. Identification of a 910-Kb region of common allelic loss in chromosome bands 16q24.1-q24.2 in human lung cancer Peer-reviewed

    M Sato, Y Mori, A Sakurada, S Fukushige, Y Ishikawa, E Tsuchiya, Y Saito, T Nukiwa, S Fujimura, A Horii

    GENES CHROMOSOMES & CANCER　22　(1)　1-8　1998/05

    ISSN： 1045-2257

  62. Infrequent genetic alterations of the PTEN gene in Japanese patients with sporadic prostate cancer Peer-reviewed

    K Orikasa, S Fukushige, S Hoshi, S Orikasa, K Kondo, Y Miyoshi, Y Kubota, A Horii

    JOURNAL OF HUMAN GENETICS　43　(4)　228-230　1998

    DOI： 10.1007/s100380050078 　

    ISSN： 1434-5161

  63. Genomic analysis of DUSP6, a dual specificity MAP kinase phosphatase, in pancreatic cancer Peer-reviewed

    T Furukawa, T Yatsuoka, EM Youssef, T Abe, T Yokoyama, S Fukushige, E Soeda, M Hoshi, Y Hayashi, M Sunamura, M Kobari, A Horii

    CYTOGENETICS AND CELL GENETICS　82　(3-4)　156-159　1998

    ISSN： 0301-0171

  64. Frequent gain of copy number on the long arm of chromosome 20 in herman pancreatic adenocarcinoma Peer-reviewed

    S Fukushige, FM Waldman, M Kimura, T Abe, T Furukawa, M Sunamura, M Kobari, A Horii

    GENES CHROMOSOMES & CANCER　19　(3)　161-169　1997/07

    DOI： 10.1002/(SICI)1098-2264(199707)19:3<161::AID-GCC5>3.0.CO;2-W 　

    ISSN： 1045-2257

  65. A novel tandem repeat sequence located on human chromosome 4p: isolation and characterization. International-journal

    M Kogi, S Fukushige, C Lefevre, S Hadano, J E Ikeda

    Genomics　42　(2)　278-83　1997/06/01

    ISSN： 0888-7543

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    In an effort to analyze the genomic region of the distal half of human chromosome 4p, to where Huntington disease and other diseases have been mapped, we have isolated the cosmid clone (CRS447) that was likely to contain a region with specific repeat sequences. Clone CRS447 was subjected to detailed analysis, including chromosome mapping, restriction mapping, and DNA sequencing. Chromosome mapping by both a human-CHO hybrid cell panel and FISH revealed that CRS447 was predominantly located in the 4p15.1-15.3 region. CRS447 was shown to consist of tandem repeats of 4.7-kb units present on chromosome 4p. A single EcoRI unit was subcloned (pRS447), and the complete sequence was determined as 4752 nucleotides. When pRS447 was used as a probe, the number of copies of this repeat per haploid genome was estimated to be 50-70. Sequence analysis revealed that it contained two internal CA repeats and one putative ORF. Database search established that this sequence was unreported. However, two homologous STS markers were found in the database. We concluded that CRS447/pRS447 is a novel tandem repeat sequence that is mainly specific to human chromosome 4p.

  66. Alternative splicing of hMSH2 in normal human tissues Peer-reviewed

    Y Mori, H Shiwaku, S Fukushige, S Wakatsuki, M Sato, T Nukiwa, A Horii

    HUMAN GENETICS　99　(5)　590-595　1997/05

    DOI： 10.1007/s004390050411 　

    ISSN： 0340-6717

  67. Alternative splicing of GTBP in normal human tissues Peer-reviewed

    Hiromi O. Shiwaku, Shigeru Wakatsuki, Yuriko Mori, Shinichi Fukushige, Akira Horii

    DNA Research　4　(5)　359-362　1997

    Publisher: Universal Academy Press Inc.

    DOI： 10.1093/dnares/4.5.359 　

    ISSN： 1340-2838

  68. Frequent gains on chromosome arms 1q and/or 8q in human endometrial cancer Peer-reviewed

    Akihiko Suzuki, Shinichi Fukushige, Satoru Nagase, Noriaki Ohuchi, Susumu Satomi, Akira Horii

    Human Genetics　100　(5-6)　629-636　1997

    DOI： 10.1007/s004390050565 　

    ISSN： 0340-6717

  69. Trapping of mammalian promoters by Cre-lox site-specific recombination. International-journal Peer-reviewed

    S Fukushige, J E Ikeda

    DNA research : an international journal for rapid publication of reports on genes and genomes　3　(2)　73-80　1996/04/30

    ISSN： 1340-2838

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    One of the challenges in human genome research is to identify the promoter sequences which play a key role in the regulation of gene expression. We report here a new promoter trapping system for use with mammalian cells comprised of the following three steps: 1) Cloning of DNA fragments into a promotertrapping vector, 2) integration of the trapping vector into a designated target in the mammalian genome using the Cre site-specific recombinase, and 3) screening of integrants for trapped promoter sequences by activation of the luciferase gene. To assess the efficiency of this system, lox trapping vectors containing sense tk promoter, antisense tk promoter, or a non-promoter sequence of the neo gene were employed. The resulting levels of luciferase activity of the site-specific integrants were measured directly. Luciferase activity of the integrants can be assayed under conventional culture conditions by simply replacing the culture medium with potassium phosphate buffer containing luciferin. Only those G418r colonies carrying the tk promoter in the normal orientation exhibited a 21-to 35-fold increase in luciferase activity over that of the other integrants. These results indicate that this system is an effective means of trapping promoter sequences from random mammalian genomic DNA fragments.

  70. Frameshift mutation at codon 642 of the hMLH1 gene in human endometrial cancer Peer-reviewed

    S Fukushige, S Wakatsuki, S Nagase, A Horii

    HUMAN MUTATION　8　(4)　394-395　1996

    DOI： 10.1002/(SICI)1098-1004(1996)8:4<394::AID-HUMU22>3.0.CO;2-X 　 10.1002/humu.1380080404 　

    ISSN： 1059-7794

  71. Genomic targeting with a positive-selection lox integration vector allows highly reproducible gene expression in mammalian cells. International-journal Peer-reviewed

    S Fukushige, B Sauer

    Proceedings of the National Academy of Sciences of the United States of America　89　(17)　7905-9　1992/09/01

    ISSN： 0027-8424

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    Stable transformants of mammalian cells from gene transfer often show extreme variability in expression of the introduced transgene. This occurs from the highly variable number of copies integrated into the genome and from position effects on gene expression due to random integration. We have eliminated both of these constraints on predictable gene expression by use of a lox recombination vector. The positive selection vector system is designed to directly select Cre-mediated DNA integration at a lox target previously placed into the genome of cultured mammalian cells. Proper targeting activates expression of a defective lox-neomycin phosphotransferase (neo) fusion gene target. With CHO cell lines containing this target, almost all of the selected transformants (54 of 56 independent G418-resistant colonies) were simple single-copy integrants of the targeting DNA. To monitor gene expression at a single chromosomal site, we used a beta-actin promoter-lacZ reporter construct. Independent G418-resistant colonies from site-specific integration of the reporter gene all showed nearly identical levels of beta-galactosidase activity when the reporter construct integrated at a particular chromosomal position. The same construct integrated at a second chromosomal position exhibited a slightly different level of activity, characteristic of that second position. These results show that Cre-mediated site-specific integration can facilitate the construction of isogenic cell lines and thereby permit reproducible gene expression in stably transformed cell lines.

  72. Construction of isogenic cell lines expressing human and rat angiotensin II AT1 receptors by Cre-mediated site-specific recombination.

    Sauer B, Baubonis W, Fukushige S, Santomenna L

    Methods: A Companion to Methods in Enzymol　4　143-149　1992

  73. TECHNICAL STUDY ON ANALYSIS OF DOUBLE STAINED HUMAN-CHROMOSOMES Peer-reviewed

    SI YOSHIMURA, M YAMAZAKI, K TAKAHARA, H SUGINO, N NAKAMURA, S FUKUSHIGE, T MUROTSU, K MATSUBARA

    FLOW CYTOMETRY AND IMAGE ANALYSIS FOR CLINICAL APPLICATIONS　933　89-94　1991

  74. Endogenous retroviral LTR DNA sequences as markers for individual human chromosomes. International-journal Peer-reviewed

    N Nakamura, H Sugino, K Takahara, C Jin, S Fukushige, K Matsubara

    Cytogenetics and cell genetics　57　(1)　18-22　1991

    ISSN： 0301-0171

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    The human genome carries multiple copies of sequences related to endogenous retroviral genomes that include long terminal repeat (LTR) sequences. We used the LTR of one such viral genome, called HERV-A, as a probe in Southern analysis to examine the distribution profiles of the hybridizing DNA in the genomes of twelve human x rodent hybrid cell lines carrying one or a few human chromosomes, and in the DNA samples prepared from six sorted, individual chromosomes. The HERV-A sequence was found to be widely distributed among different chromosomes and the Southern patterns for chromosomes 5, the X, and the Y, each obtained in duplicate from independently prepared cell lines or sorted chromosomes, were matched. Chromosome-specific Southern profiles can be used to monitor chromosomes in hybrid cells or to characterize chromosome aberrations, such as deletions.

  75. Separation of DNA restriction fragments by high-performance ion-exchange chromatography on a non-porous ion exchanger. International-journal Peer-reviewed

    Y Kato, Y Yamasaki, A Onaka, T Kitamura, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of chromatography　478　(1)　264-8　1989/09/08

  76. Two erbA homologs encoding proteins with different T3 binding capacities are transcribed from opposite DNA strands of the same genetic locus. International-journal Peer-reviewed

    N Miyajima, R Horiuchi, Y Shibuya, S Fukushige, K Matsubara, K Toyoshima, T Yamamoto

    Cell　57　(1)　31-9　1989/04/07

    ISSN： 0092-8674

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    Two erbA homologs, termed ear-1 and ear-7, are present in the human genome on chromosome 17. The two genes reside in the same genetic locus with overlapping exons and are transcribed from opposite DNA strands. In addition, the ear-7 mRNA is alternatively spliced to generate two protein isoforms, namely the ear71 and ear72 proteins. Nucleotide sequence analysis predicts that the ear71 protein is a human counterpart of the chicken c-erbA protein, a molecule closely related or identical to thyroid hormone receptor. Indeed, Scatchard analysis of proteins synthesized in vitro indicated very high affinity binding of T3 to the ear71 protein but not to the ear72 protein. Interestingly, the ear-1 gene product showed low, but appreciable, binding to T3, although its authentic ligand remains to be clarified.

  77. Assignment of human pepsinogen C (PGC) gene to chromosome 6. International-journal Peer-reviewed

    K Takahara, S Fukushige, T Murotsu, Y Ichihara, T Hayano, T Ishihara, K Takahashi, K Matsubara

    Cytogenetics and cell genetics　52　(1-2)　100-1　1989

    ISSN： 0301-0171

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    cDNA of rat pepsinogen C (PGC) hybridizes to, among others, a 3.2-kb band in Southern blot analysis of BamHI-cleaved human genomic DNA. This property was employed to localize the human PGC gene. Use of flow-sorted human chromosomes and 12 human x mouse somatic cell hybrid lines demonstrated that the gene is located on chromosome 6.

  78. Identification of two novel members of erbA superfamily by molecular cloning: the gene products of the two are highly related to each other. International-journal Peer-reviewed

    N Miyajima, Y Kadowaki, S Fukushige, S Shimizu, K Semba, Y Yamanashi, K Matsubara, K Toyoshima, T Yamamoto

    Nucleic acids research　16　(23)　11057-74　1988/12/09

    ISSN： 0305-1048

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    Two v-erbA-related genes, named ear-2 and ear-3, have been identified in the human genome and characterized by cDNA cloning. These genes are predicted to encode proteins that are very similar in primary structure to receptors for steroid hormones or thyroid hormone (T3). In addition, amino acid sequences of the ear-2 and ear-3 gene products are very similar each other especially at the DNA binding domain (86% homology) and at the putative ligand binding domain (76% homology). Northern hybridization with ear DNA probes of RNAs from various tissues of a human fetus reveals that the expression of ear-2 is high in the liver whereas the expression of ear-3 is relatively ubiquitous. Hybridization analysis of DNAs from sorted chromosomes shows that the ear-2 gene is located on chromosome 19 and ear-3 on chromosome 5, indicating that the two genes are clearly different from each other.

  79. Separation of oligonucleotides by high-performance ion-exchange chromatography on a non-porous ion exchanger. International-journal Peer-reviewed

    Y Kato, T Kitamura, A Mitsui, Y Yamasaki, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of chromatography　447　(1)　212-20　1988/08/05

  80. Nucleotide sequence and chromosomal mapping of the human c-yes-2 gene. Peer-reviewed

    K Semba, M Nishizawa, H Satoh, S Fukushige, M C Yoshida, M Sasaki, K Matsubara, T Yamamoto, K Toyoshima

    Japanese journal of cancer research : Gann　79　(6)　710-7　1988/06

    ISSN： 0910-5050

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    We molecularly characterized the second gene, c-yes-2, of two copies of yes-related genes which we previously found to contain in the human genome. First, nucleotide sequence analysis revealed that the c-yes-2 gene is a pseudogene of the c-yes-1 gene. Second, by using two independent methods, hybridization of both DNAs from sorted chromosomes and metaphase spreads with c-yes-2 DNA, we assigned the c-yes-2 gene to chromosome 22q11.2. This chromosomal localization is consistent with that given in our previous report. The failure of proper mapping in our experiment might have been caused by instability of hybrid cell clones.

  81. Primary structure of human pancreatic secretory trypsin inhibitor (PSTI) gene. Peer-reviewed

    Horii A, Kobayashi T, Tomita N, Yamamoto T, Fukushige S, Murotsu T, Ogawa M, Mori T, Matsubara K

    Biochem Biophys Res Commun　149　(2)　635-641　1987/12/16

    DOI： 10.1016/0006-291X(87)90415-3 　

  82. Chromosomal translocation and inverted duplication associated with integrated hepatitis B virus in hepatocellular carcinomas. International-journal Peer-reviewed

    T Tokino, S Fukushige, T Nakamura, T Nagaya, T Murotsu, K Shiga, N Aoki, K Matsubara

    Journal of virology　61　(12)　3848-54　1987/12

    ISSN： 0022-538X

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    Integrated hepatitis B virus (HBV) DNA is found in hepatocellular carcinomas which develop in HBV carriers. Presented here are the results of analyses of four integrants that show chromosomal rearrangements associated with the integrated HBV DNA. Two clones (p4 and C15) were found to have large inverted repeating structures, each consisting of HBV genome along with flanking cellular sequences. The structure must have arisen by duplication of the primary integrant, including the flanking cellular DNA, followed by recombination within the viral DNA. One of the two viral arms in each clone joins to the other viral arm at the "cohesive end region." Two clones (DA2-2 and DA2-6) were found to have integrated HBV sequences, each flanked by cellular DNAs from different chromosomes (chromosome X joined to 17 and chromosome 5 joined to 9). They must be the products of cellular DNA translocations using the integrated HBV DNA as the switch point. The viral DNA in each clone is a continuous stretch of a single virus genome with one end in the cohesive end region. These complex structures seem to have been produced by activation of the cohesive end of an integrant viral genome, followed by its recombination with another chromosomal DNA.

  83. Human gene coding for granulocyte-colony stimulating factor is assigned to the q21-q22 region of chromosome 17. International-journal Peer-reviewed

    N Kanda, S Fukushige, T Murotsu, M C Yoshida, M Tsuchiya, S Asano, Y Kaziro, S Nagata

    Somatic cell and molecular genetics　13　(6)　679-84　1987/11

    ISSN： 0740-7750

    More details Close

    Granulocyte-colony stimulating factor (G-CSF) is a member of colony stimulating factors which regulate the proliferation and differentiation of hematopoietic progenitor cells. A full-length cDNA clone coding human G-CSF was used as a hybridization probe to detect homologous sequence in human-mouse somatic cell hybrids, flow-sorted human chromosomes, and in situ human metaphase chromosomes. The results indicate that the gene encoding human G-CSF is on the q21-q22 region of chromosome 17, which is involved in translocation of t(15;17) (q23;21) in human acute promyelocytic leukemia.

  84. Isolation and characterization of the active cDNA of the human cell cycle gene (RCC1) involved in the regulation of onset of chromosome condensation. International-journal Peer-reviewed

    M Ohtsubo, R Kai, N Furuno, T Sekiguchi, M Sekiguchi, H Hayashida, K Kuma, T Miyata, S Fukushige, T Murotsu

    Genes & development　1　(6)　585-93　1987/08

    ISSN： 0890-9369

    More details Close

    The human RCC1 gene was cloned after DNA-mediated gene transfer into the tsBN2 cell line, which shows premature chromosome condensation at nonpermissive temperatures (39.5-40 degrees C). This gene codes for a 2.5-kb poly(A)+ RNA that is well conserved in hamsters and humans. We isolated 15 cDNA clones from the Okayama-Berg human cDNA library, and found two that can complement the tsBN2 mutation with an efficiency comparable to that of the genomic DNA clone. The base sequences of these two active cDNA clones differ at the 5' proximal end, yet both have a common open reading frame, encoding a protein of 421 amino acids with a calculated molecular weight of 44,847 and with seven homologous repeated domains of about 60 amino acids. This human RCC1 gene was located to human chromosome 1 using sorted chromosomal fractions.

  85. Molecular gene mapping of human aldolase A (ALDOA) gene to chromosome 16. International-journal Peer-reviewed

    A Kukita, M C Yoshida, S Fukushige, M Sakakibara, K Joh, T Mukai, K Hori

    Human genetics　76　(1)　20-6　1987/05

    ISSN： 0340-6717

    More details Close

    Mapping of human aldolase A (ALDOA) gene was performed by molecular hybridization techniques using a panel of human-mouse cell hybrids and sorted fractions of human metaphase chromosomes besides in situ hybridization. For the purpose, three kinds of DNA probes derived from the coding region (probe-1), the 3' noncoding region (probe-2), and the coding and 3' noncoding regions (probe-3) of human aldolase A cDNA clone, pHAAL116-3, were selectively employed. The results of RNA and DNA blot analyses indicated that the human ALDOA gene is located on chromosome 16. The in situ hybridization experiment also indicated that the ALDOA gene was localized to 16q22-q24.

  86. Purification of plasmid by high-performance gel filtration. Peer-reviewed

    Yamasaki Y, Kato Y, Murotsu T, Fukushige S, Matsubara K

    J High Resolut Chromatogr Chromatogr Commun　10　45-46　1987

  87. The yes-related cellular gene lyn encodes a possible tyrosine kinase similar to p56lck. International-journal Peer-reviewed

    Y Yamanashi, S Fukushige, K Semba, J Sukegawa, N Miyajima, K Matsubara, T Yamamoto, K Toyoshima

    Molecular and cellular biology　7　(1)　237-43　1987/01

    ISSN： 0270-7306

    More details Close

    With v-yes DNA as the probe, a human cDNA library made from placental RNA was screened under relaxed conditions, and DNA clones derived from a novel genetic locus, termed lyn, were obtained. Nucleotide sequencing revealed that lyn could encode a novel tyrosine kinase that was very similar to mouse T-lymphocyte-specific tyrosine kinase p56lck and the v-yes protein as well as to the gene products of v-fgr and v-src. Northern hybridization analysis revealed that a 3.2-kilobase lyn mRNA was expressed in a variety of tissues of the human fetus. The pattern of lyn mRNA expression was different from those of related genes, such as yes and syn. Hybridization analysis of DNA from sorted chromosomes showed that the lyn gene is located on human chromosome 8 q13-qter.

  88. Molecular cloning of an oncogene from a human hepatocellular carcinoma. International-journal Peer-reviewed

    T Ochiya, A Fujiyama, S Fukushige, I Hatada, K Matsubara

    Proceedings of the National Academy of Sciences of the United States of America　83　(14)　4993-7　1986/07

    ISSN： 0027-8424

    More details Close

    A transforming DNA, named lca (for liver cancer), was obtained from a primary human hepatocellular carcinoma (HCC) in transformation assays using NIH 3T3 cells and a calcium phosphate coprecipitation method. High molecular weight DNA obtained from the HCC tissue was employed for this purpose. This transforming DNA had a linkage to the Alu sequence and was cloned in lambda phage for further studies. Restriction enzyme analyses showed that the minimal size of the lca transforming DNA is about 10 kilobase pairs and that its cleavage profiles are different from those of any one of the previously reported human transforming genes or retroviral oncogenes. No cross-hybridization was observed between these genes and the lca DNA. Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids indicated that the lca DNA is located on human chromosome 2. An independently obtained transforming DNA from another HCC exhibited identical restriction enzyme cleavage profiles. Thus, lca DNA is likely to represent a commonly encountered transforming DNA in HCC.

  89. Localization of a novel v-erbB-related gene, c-erbB-2, on human chromosome 17 and its amplification in a gastric cancer cell line. International-journal Peer-reviewed

    S Fukushige, K Matsubara, M Yoshida, M Sasaki, T Suzuki, K Semba, K Toyoshima, T Yamamoto

    Molecular and cellular biology　6　(3)　955-8　1986/03

    ISSN： 0270-7306

    More details Close

    The c-erbB-2 gene is a v-erbB-related proto-oncogene which is distinct from the gene encoding the epidermal growth factor receptor. By using two independent methods, hybridization of both sorted chromosomes and metaphase spreads with cloned c-erbB-2 DNA, we mapped the c-erbB-2 locus on human chromosome 17 at q21, a specific breakpoint observed in a translocation associated with acute promyelocytic leukemia. Furthermore, we observed amplification and elevated expression of the c-erbB-2 gene in the MKN-7 gastric cancer cell line. These data suggest possible involvement of the c-erbB-2 gene in human cancer.

  90. Chromosomal assignment of human genes for gastrin, thyrotropin (TSH)-beta subunit and C-erbB-2 by chromosome sorting combined with velocity sedimentation and Southern hybridization. International-journal Peer-reviewed

    S Fukushige, T Murotsu, K Matsubara

    Biochemical and biophysical research communications　134　(2)　477-83　1986/01/29

    ISSN： 0006-291X

    More details Close

    Human genes for gastrin, thyrotropin (THS)-beta subunit and c-erbB-2 were assigned to specific chromosomes using a single-laser cell sorter. For this purpose, condensed human chromosomes prepared from a karyotypically normal lymphoblastoid cell line were preliminarily fractionated by velocity sedimentation, and then sorted using a fluorescence-activated cell sorter. DNA was then extracted from the chromosomes, cleaved by restriction enzymes, and subjected to Southern hybridization using gene-specific radioactive probes. When the assignment of specific chromosomes was not possible due to chromosomal size overlapping, sorted chromosomes from cell lines carrying chromosomal translocation or from hybrid cells carrying known human chromosomes were used in addition. The results indicate that human genes for gastrin, TSH-beta, and c-erbB-2 are located on chromosomes 17, 1 and 17, respectively.

  91. Structure of human cholecystokinin gene and its chromosomal location. International-journal Peer-reviewed

    Y Takahashi, S Fukushige, T Murotsu, K Matsubara

    Gene　50　(1-3)　353-60　1986

    ISSN： 0378-1119

    More details Close

    We have determined the entire structure of human cholecystokinin (CCK) gene, which is 7 kb in size and separated into three exons. S1 endonuclease analysis has shown two putative transcription initiation sites that are preceded by 'TATA' equivalent sequences located 39 bp and 35 bp upstream from these sites. The promoter region contains five 'G-C box'-like sequences, which are believed to be sp 1-binding sites. By chromosome sorting in combination with velocity sedimentation and Southern hybridization, the human cck gene was mapped on the short arm of human chromosome 3.

  92. Separation of large DNA restriction fragments by high-performance gel filtration on TSKgel DNA-PW. International-journal Peer-reviewed

    Y Kato, Y Yamasaki, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of chromatography　320　(2)　440-4　1985/03/01

  93. A new packing for separation of DNA restriction fragments by high performance liquid chromatography. International-journal Peer-reviewed

    Y Kato, M Sasaki, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of biochemistry　95　(1)　83-6　1984/01

    ISSN： 0021-924X

    More details Close

    TSK-GEL SW was found to be useful as a packing in high performance liquid chromatography for the separation of double-stranded DNA restriction fragments. DNA fragments smaller than 300 base pairs were separated as discrete peaks depending solely upon difference in chain length. The recovery of DNA fragments was higher than 90%.

  94. Operational variables in high-performance gel filtration of DNA fragments and RNAs. International-journal Peer-reviewed

    Y Kato, M Sasaki, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of chromatography　266　341-9　1983/08/26

    More details Close

    Double-stranded DNA fragments and ribosomal and transfer RNAs were measured by high-performance gel filtration on TSK-GEL G2000SW, G3000SW, G4000SW and G5000PW columns to investigate the separation range and resolution of these columns and the effects of eluent ionic strength and flow-rate on retention and resolution. These columns could separate double-stranded DNA fragments up to ca 1 X 10(6) and rRNAs up to ca 5 X 10(6) daltons in molecular weight. However, it was found that the selection of the column is very important to achieve optimum separation, depending on the molecular weight of the sample. Elution is delayed as the eluent ionic strength is increased. An eluent ionic strength of 0.3-0.5 seemed appropriate in most cases. Resolution is greatly increased as the flow-rate is decreased.

  95. Separation of DNA restriction fragments by high-performance ion-exchange chromatography. International-journal Peer-reviewed

    Y Kato, M Sasaki, T Hashimoto, T Murotsu, S Fukushige, K Matsubara

    Journal of chromatography　265　(2)　342-6　1983/08/19

  96. Loading capacity in high performance gel filtration of nucleic-acid on TSK-GEL SW. Peer-reviewed

    Kato Y, Hashimoto T, Murotsu T, Fukushige S, Matsubara K

    J High Resolut Chromatogr Chromatogr Commun　6　626-626　1983

Show all ︎Show first 5

Misc. 34

  1. Cytidine deaminase plays a critical role in acquisition of gemcitabine resistance in a pancreatic cancer cell line

    日吉貴子, 斉木由利子, 佐々木彰之, 廣田嵩人, 平山明由, 曽我朋義, 福重真一, 堀井明

    日本癌学会学術総会抄録集(Web)　78th　2019

  2. 前立腺腫瘍形成においてPYCARD遺伝子プロモーターのメチル化が高頻度に発生する

    福重 真一, 宮内 隼弥, 大久保 鉄平, 齋木 由利子, 高橋 正博, 三塚 浩二, 荒井 陽一, 堀井 明

    日本癌学会総会記事　76回　P-1153　2017/09

    Publisher: 日本癌学会

    ISSN： 0546-0476

  3. Targeted TET oxidase activity through methyl-CpG binding domain extensively suppresses cancer cell proliferation Peer-reviewed

    Shinichi Fukushige, Yasuhiko Mizuguchi, Kanchan Chakma, Yuriko Saiki, Akira Horii

    CANCER RESEARCH　76　2522-2533　2016/07

    DOI： 10.1158/1538-7445.AM2016-4432 　

    ISSN： 0008-5472

    eISSN： 1538-7445

  4. TET oxidase activity accumulated on methyl-CpG sites extensively upregulates methylated genes through DNA demethylation

    Shinichi Fukushige, Yasuhiko Mizuguchi, Akira Horii

    CANCER RESEARCH　74　(19)　2014/10

    DOI： 10.1158/1538-7445.AM2014-396 　

    ISSN： 0008-5472

    eISSN： 1538-7445

  5. IFI27 and NOV, downstream regulated genes by S100A4, are playing important roles in pancreatic carcinogenesis

    Na Chen, Yuriko Saiki, Hitoshi Sekine, Makoto Sunamura, Shinichi Fukushige, Fuyuhiko Motoi, Shinichi Egawa, Michiaki Unno, Akira Horii

    CANCER RESEARCH　74　(19)　2014/10

    DOI： 10.1158/1538-7445.AM2014-4993 　

    ISSN： 0008-5472

    eISSN： 1538-7445

  6. Highlights from the latest articles in epigenomics. International-journal Peer-reviewed

    Shinichi Fukushige, Akira Horii

    Epigenomics　6　(2)　171-3　2014/04

    DOI： 10.2217/epi.14.2 　

  7. Rosvastatin Ameliorates Lung Ischemic-Reperfusion Injury via eNOS Upregulation and Inhibition of Macrophage Recruitment in Pulmonary Hypertensive Rat

    Satoshi Matsuo, Yuriko Saiki, Koki Ito, Yukihiro Hayatsu, Yusuke Suzuki, Shinichi Fukushige, Shunsuke Kawamoto, Akira Horii, Yoshikatsu Saiki

    CIRCULATION　128　(22)　2013/11

    ISSN： 0009-7322

    eISSN： 1524-4539

  8. ゲムシタビン耐性ヒト癌細胞株におけるdeoxycytidine kinaseの不活化(Deoxycytidine kinase is frequently inactivated in acquired gemcitabine-resistant human cancer cells)

    齋木 由利子, 吉野 優樹, 福重 真一, 砂村 真琴, 石田 晶玄, 元井 冬彦, 江川 新一, 海野 倫明, 堀井 明

    日本癌学会総会記事　71回　237-237　2012/08

    Publisher: 日本癌学会

    ISSN： 0546-0476

  9. 大腸癌における新規癌抑制遺伝子NDRG2のエピジェネティックな発癌制御機構

    山村 明寛, 堀井 明, 三浦 康, 唐沢 秀明, 阿部 佳子, 内藤 剛, 小川 仁, 木内 誠, 矢崎 伸樹, 羽根田 祥, 渡辺 和宏, 大沼 忍, 佐瀬 友彦, 鈴木 秀幸, 斉木 由利子, 福重 真一, 柴田 近, 佐々木 巖

    日本大腸肛門病学会雑誌　64　(8)　537-537　2011/08

    Publisher: (一社)日本大腸肛門病学会

    ISSN： 0047-1801

  10. Identification of novel targets for aberrant methylation in pancreatic cancer by a newly developed method "methyl-CpG targeted transcriptional activation (MeTA)"

    Shinichi Fukushige, Hideyuki Shimizu, Yuko Sato, Akira Horii

    CANCER RESEARCH　70　2010/04

    DOI： 10.1158/1538-7445.AM10-179 　

    ISSN： 0008-5472

    eISSN： 1538-7445

  11. Expression of the N-myc downstream-regulated gene 2 (NDRG2) is frequently suppressed by promoter hypermethylation in human gastrointestinal and pancreatic cancers

    Akihiro Yamamura, Koh Miura, Hideaki Karasawa, Keiko Abe, Yasuhiko Mizuguchi, Guo Jin, Zhaodi Gu, Shinichi Fukushige, Naoyuki Kaneko, Makoto Kinouchi, Toshinori Ando, Nobuki Yazaki, Naoki Tanaka, Tomohiko Sase, Chikashi Shibata, Iwao Sasaki, Akira Horii

    CANCER RESEARCH　70　2010/04

    DOI： 10.1158/1538-7445.AM10-4933 　

    ISSN： 0008-5472

    eISSN： 1538-7445

  12. 癌抑制遺伝子NDRG2のエピジェネティックサイレンシングは消化器発癌を進行させる

    山村 明寛, 三浦 康, 木内 誠, 安藤 敏典, 矢崎 伸樹, 田中 直樹, 唐澤 秀明, 佐瀬 友彦, 木村 俊一, 福重 真一, Gu Zhaodi, 嶋 健太郎, 柴田 近, 堀井 明, 佐々木 巖

    日本外科学会雑誌　111　(臨増2)　384-384　2010/03

    Publisher: (一社)日本外科学会

    ISSN： 0301-4894

  13. 消化器癌においてNDRG2はプロモーターのメチル化によって発現抑制される(Promoter hypermethylation and reduced expression of NDRG2 in gastrointestinal cancers)

    山村 明寛, 三浦 康, 木内 誠, 安藤 敏典, 矢崎 伸樹, 田中 直樹, 唐澤 秀明, 佐瀬 友彦, 福重 真一, Gu Zhaodi, 柴田 近, 佐々木 巖, 堀井 明寛

    日本癌学会総会記事　68回　173-173　2009/08

    Publisher: 日本癌学会

    ISSN： 0546-0476

  14. Whole Genome Amplificationを利用した膵癌の遺伝子診断 Peer-reviewed

    阿部忠義, 砂村眞琴, 江川新一, 福山尚治, 元井冬彦, 福重真一, 堀井明, 海野倫明

    日本消化器外科学会雑誌　39　(7)　1180-1180　2006/07

  15. PanIN病変,膵癌のWhole Genome Amplificationを用いたゲノム解析 Peer-reviewed

    阿部忠義, 砂村眞琴, 江川新一, 福山尚治, 元井冬彦, 福重真一, 堀井明, 海野倫明

    膵臓　21　(3)　249-249　2006/06

  16. Fluorescence in situ hybridization analysis of breast cancer: Positive association between loss of 17p13 and HER2 overexpression Peer-reviewed

    Mitsue Oguma, Takuya Moriya, Shinichi Fukushige, Takanori Ishida, Akira Horii, Noriaki Ohuchi

    FUTURE MEDICAL ENGINEERING BASED ON BIONANOTECHNOLOGY, PROCEEDINGS　489-+　2006

    DOI： 10.1142/9781860948800_0053 　

  17. 膵液中の細胞を用いた膵癌の遺伝子診断 Peer-reviewed

    砂村眞琴, 福重真一, 堀井明, 阿部忠義, 松野正紀

    コンセンサス癌治療　2　(1)　46-49　2003/02

  18. Isolation and characterization of the human gene homologous to the Drosophila headcase (hdc) gene in chromosome bands 6q23-q24, a region of common deletion in human pancreatic cancer Peer-reviewed

    Naohiko Makino, Takashi Yamato, Hiroko Inoue, Toru Furukawa, Tadayoshi Abe, Tadaaki Yokoyama, Toshimasa Yatsuoka, Shinichi Fukushige, Seiichi Orikasa, Tsuneo Takahashi, Akira Horii

    Mitochondrial DNA　11　(6)　547-553　2001

    DOI： 10.3109/10425170109041340 　

    ISSN： 1940-1736 1940-1744

  19. Allelic imbalanceを指標にした膵癌の存在診断・悪性度診断 Invited

    八岡利昌, 福重真一, 堀井明

    消化器科　30　(1)　98-105　2000/01

  20. Early diagnosis of pancreatic ductal tumors by using cells from pancreatic juice

    FUKUSHIGE Shinichi, HORII Akira

    9　(1)　53-58　1999/06/25

    ISSN： 0916-6920

  21. Comparative genomic hybridization(CGH) 膵液中の細胞を用いた膵腫瘍の早期診断 Invited

    福重真一, 堀井明

    Cytometry Research　9　(1)　53-58　1999/06

  22. WS1a-3 第18番染色体長腕の遺伝子異常は膵癌発生の初期変化である : 膵液中の細胞のFISH法による検討

    福重 真一, 砂村 眞琴, 小針 雅男, 松野 正紀, 堀井 明

    日本外科学会雑誌　100　54-54　1999/02/10

    Publisher: 一般社団法人日本外科学会

    ISSN： 0301-4894

  23. すいがんで高頻度に欠失している第12番染色体長腕の解析

    古川徹, 八岡利昌, YOUSSEF E M, 福重真一, 砂村真琴, 堀井明

    日本癌学会総会記事　58th　1999

    ISSN： 0546-0476

  24. すい液中細胞利用によるすい腫ようの新たな遺伝子診断法の開発 染色体・遺伝子異常を指標としたすい腫ようの個性化の試み

    八岡利昌, 福重真一, 古川徹, 堀井明

    日本人類遺伝学会大会プログラム・抄録集　44th　1999

  25. Allelic imbalanceを指標にしたすい癌の存在診断および悪性度診断の検討

    八岡利昌, 古川徹, 福重真一, 木村光宏, 阿部忠義, 砂村真琴, 武田和憲, 松野正紀, 堀井明

    日本癌学会総会記事　58th　1999

    ISSN： 0546-0476

  26. Analysis of the protein interactions between hPMS2 family and hMLH1 and other proteins

    FUKUSHIGE Shinichi, HORII Akira

    21　373-373　1998/12/01

  27. 膵腫瘍の診断において膵液中の細胞を用いたFISH法は有用である

    福重 真一, 古川 徹, 佐藤 賢一, 砂村 眞琴, 小針 雅男, 堀井 明

    日本癌学会総会記事　57　713-713　1998/08

  28. W2-7 遺伝子解析に基づいた膵癌に対する新たな診療体系の確立(第52回日本消化器外科学会総会)

    砂村 眞琴, 木村 光宏, 阿部 忠義, 丁 良浩, 福山 尚治, 江川 新一, 小針 雅男, 堀井 明, 福重 真一, 古川 徹, 八岡 利昌, 横山 忠明, 濱田 洋文, 元井 冬彦, 吉田 陽子, 松野 正紀

    日本消化器外科学会雑誌　31　(6)　1314-1314　1998/06/01

    Publisher: 一般社団法人日本消化器外科学会

    ISSN： 0386-9768

  29. 膵癌のCGH解析 (組織培養工学)

    福重真一, 堀井明

    組織培養工学　23　(9)　339-344　1997/08

  30. Genetic instabilityと発癌 マイクロサテライト不安定性と発癌 (血液・腫瘍科)

    福重真一, 堀井明

    血液・腫瘍科　34　(6)　431-438　1997/06

  31. [Disruption of mismatch repair system in human cancers].

    S Fukushige, A Horii

    Nihon rinsho. Japanese journal of clinical medicine　54　(4)　1002-7　1996/04

    ISSN： 0047-1852

    More details Close

    It is known that transformation of normal cells to cancer cells is caused by the accumulation of successive mutations in oncogenes and/or tumor suppressor genes. Since four DNA mismatch repair genes (hMSH2, hMLH1, hPMS1 and hPMS2) have been identified as the cause of hereditary nonpolyposis colorectal cancer (HNPCC), the role of defective mismatch repair system in the development of sporadic cancers with microsatellite instability has also been discussed. Defects in mismatch repair genes would contribute to mutations in genes, including oncogenes and tumor suppressor genes, at an increased rate. Furthermore, recent investigations suggested that this mechanism was also involved in the development of multiple primary cancers as well.

  32. DNAミスマッチ修復遺伝子異常とreplication error(genomic instability) 発がんにおけるDNAミスマッチ修復遺伝子の異常の関与 (日本臨床)

    福重真一, 堀井明

    日本臨床　54　(4)　1002-1007　1996/04

  33. Genetic alterations in human pancreatic cancer Peer-reviewed

    A Horii, M Kimura, T Abe, S Fukushige, T Furukawa, M Sunamura, M Kobari, S Matsuno

    RECENT ADVANCES IN GASTROENTEROLOGICAL CARCINOGENESIS I　231-238　1996

  34. A novel oncogene from a human hepatocellular carcinoma. International-journal

    K Matsubara, T Ochiya, I Hatada, M Shiozawa, S Fukushige, M Araki, A Fujiyama, T Tsurimoto

    Princess Takamatsu symposia　17　133-41　1986

    More details Close

    Primary human hepatocellular carcinomas (HCC) were surveyed for oncogenes by transformation assays using NIH 3T3 cells and calcium phosphate coprecipitation method. One new transforming DNA, called lca (for liver cancer) was obtained, and its properties were studied in detail. The lca DNA, localized in a 10.5 kb DNA fragment, was assigned to human chromosome 2. It showed identical restriction enzyme cleavage profiles as its counterpart DNA obtained from normal tissue, indicating that there is no extensive DNA rearrangement associated with activation process of the lca DNA. The normal counterpart DNA shows no transforming activity. Recombinant genes of the laca and its normal counterpart made in vitro have allowed localization of the site of "activation" to a limited region of the 10.5 kb fragment. An independently obtained transforming DNA from another HCC exhibited identical restriction enzyme cleavage profiles. Thus, lca DNA is likely to represent a commonly encountered transforming DNA in HCC.

Show all ︎Show first 5

Books and Other Publications 2

  1. Biomarkers in Cancer

    Fukushige S, Horii A

    Springer　2015

    ISBN: 9789400777446

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    DOI: 10.1007/978-94-007-7744-6_45-1

  2. Cytochromes P450, Biochemistry, Biophysics and Drug Metabolism. 13th International Conference on Cytochromes P450.

    Kikuchi H. Fukushige, S. Shibazaki, M. Sohel A, Takeuchi T

    Monduzzi Editore, Bologna (Italy)　2003/04

Presentations 60

  1. 膵癌細胞株においてシチジンデアミナーゼの発現増加はゲムシタビン耐性獲得に関与する

    日吉貴子、齋木由利子、佐々木彰之、廣田嵩人、平山明由、曽我朋義、福重真一、堀井 明

    第78回日本癌学会学術総会　2019/09

  2. IRX1の異常メチル化は前立腺癌に高頻度で発生し、増殖を活性化する

    高橋正博、三塚浩二、伊藤明宏、堀井明、福重真一.

    第78回日本癌学会学術総会　2019/09

  3. LHX6はエピジェネティックに不活性化される膵癌の癌抑制遺伝子候補である

    ヤクブアブドゥリシディ、堀井明、元井冬彦、海野倫明、福重真一

    第78回日本癌学会学術総会　2019/09

  4. PYCARDは前立腺癌において異常にメチル化し、発現低下している

    福重真一、宮内隼弥、高橋正博、齋木由利子、大久保鉄平、三塚浩二、荒井陽一、堀井明

    第78回日本癌学会学術総会　2019/09

  5. アポトーシス誘導遺伝子PYCARDは前立腺癌でDNA高度メチル化により発現抑制される

    宮内隼弥、高橋正博、大久保鉄平、齋木由利子、三塚浩二、荒井陽一、堀井明、福重真一.

    第108回日本病理学会総会　2019/05

  6. DNA hypermethylation of IRX4 is a frequent event that may confer growth advantage to pancreatic cancer cells.

    Chakma K, Gu Z, Motoi F, Unno M, Horii A, Fukushige S

    108th American Association for Cancer Research Annual Meeting, Atlanta, GA, USA, 2019.　2019/03

  7. Relationship between expression of S100A10 and prognosis in lung squamous carcinoma.

    佐藤公昭、齋木由利子、新井一守、石沢興太、福重真一、青木献広、櫻田晃、岡田克典、堀井明

    第77回日本癌学会学術総会　2018/09

  8. ZNF750 gene promoter is aberrantly methylated in prostate cancer.

    高橋正博、三塚浩二、齋木由利子、堀井明、荒井陽一、福重真一.

    第77回日本癌学会学術総会　2018/09

  9. IRX4, a hypermethylated gene in pancreatic cancer, regulates expression of a subset of cancer-related genes.

    2018/09

  10. Methylation of the PYCARD gene promoter is a highly frequent event in prostate tumorigenesis.

    福重真一、宮内隼弥、大久保鉄平、齋木由利子、高橋正博、三塚浩二、荒井陽一、堀井明

    第76回日本癌学会学術総会　2017/09

  11. Methylation-mediated silenced PYCARD plays a key role in human prostate cancer.

    Fukushige S, Miyauchi T, Okubo T, Mitsuzuka K, Arai Y, Horii A:

    108th American Association for Cancer Research Annual Meeting, Washington DC, USA, 2017.　2017

  12. Methylation-mediated silenced PYCARD plays a key role in human prostate cancer. International-presentation

    The 2017 Japan-NIH Joint Symposium, Sendai, Japan,　2017/02/15

  13. DNA hypermethylation of IRX4 is a frequent event that may confer growth advantage to pancreatic cancer cells.

    福重真一、チャクマ・カンチャン、元井冬彦、海野倫明、堀井明.

    第75回日本癌学会学術総会　2016/10

  14. Targeted TET oxidase activity through methyl-CpG binding domain extensively suppresses cancer cell proliferation.

    Fukushige S, Mizuguchi Y, Chakma K, Saiki Y, Horii A

    107th American Association for Cancer Research Annual Meeting, New Orleans, LA, USA, 2016.　2016/04

  15. Targeted TET oxidase activity through methyl-CpG binding domain extensively suppresses cancer cell proliferation. International-presentation

    Fukushige S, Mizuguchi Y, Chakma K, Saiki Y, Horii A:

    13th International Congress of Human Genetics, Kyoto, Japan, 2016　2016/04

  16. Methyl-CpG targeted transcriptional activation (MeTA) induces apoptosis in LNCaP human prostate cancer cells by reactivating hypermethylated genes. International-presentation

    Miyauchi T, Horii A, Fukushige S

    The 10th AACR/JCA Joint Conference,“Breakthroughs in Cancer Research: From Biology to Therapeutics”, Maui, HI, USA, 2016.　2016/02

  17. DNMT-independent reactivation of hypermethylated genes induces growth suppression and apoptosis in LNCaP cells

    福重真一, 宮内隼也, 堀井明

    第74回日本癌学会学術総会　2015/10/08

  18. Molecular characterization of oncogenic properties of S100A4 in pancreatic and lung cancers, and identification of PRDM2 and IFI27 as the candidate downstream genes International-presentation

    Akira Horii, Na Chen, Hitoshi Sekine, Nobukazu Tsukamoto, Takahiro Tabata, Yuriko Saiki, Shinichi Fukushige, Makoto Sunamura

    American Society of Human Genetics Annual Meeting 2014　2014/10/18

  19. Methyl-CpG targeted DNA demethylation by TET hydroxylase activity suppresses the growth of cancer cells

    福重真一, 齋木由利子, 堀井明

    第73回日本癌学会学術総会　2014/09/25

  20. S100A4 is frequently overexpressed in lung cancer cells and promotes cell motility

    陳娜, 齋木由利子, 砂村真琴, 福重真一, 堀井明

    第73回日本癌学会学術総会　2014/09/25

  21. TET oxidase accumulated on methyl-CpG sites extensively upregulates methylated genes through DNA demethylation International-presentation

    Shinichi Fukushige, Yasuhiko Mizuguchi, Akira Horii

    105th American Association for Cancer Research Annual Meeting　2014/04/05

  22. IFI27 and NOV, downstream regulated genes by S100A4, are playing important roles in pancreatic carcinogenesis International-presentation

    Na Chen, Yuriko Saiki, Hitoshi Sekine, Makoto Sunamura, Shinichi Fukushige, Fuyuhiko Motoi, Shinichi Egawa, Michiaki Unno, Akira Horii

    105th American Association for Cancer Research Annual Meeting　2014/04/05

  23. TET1蛋白のヒドロキシラーゼ活性はエピジェネティックに転写抑制された遺伝子の再活性化を引き起こす

    福重真一, 水口康彦, 堀井明

    第72回 日本癌学会学術総会　2013/10/03

  24. S100A4の下流で制御されるIFI27、NOVの膵癌の発生、進展における役割の解析

    陳娜, 齋木由利子, 砂村真琴, 福重真一, 元井冬彦, 江川新一, 海野倫明, 堀井明

    第72回 日本癌学会学術総会　2013/10/03

  25. MeTA-array, a useful method to identify genes transcriptionally silenced by tumor-specific hypermethylation in cancer. International-presentation

    Fukushige S, Horii A

    104th American Association for Cancer Research Annual Meeting　2013/04/06

  26. ヒト癌において腫瘍特異的な高度メチル化により転写抑制された遺伝子のMeTA-arrayによる効率的探索法の開発

    福重真一, 堀井明

    第７１回日本癌学会学術総会　2012/09/19

  27. ゲムシタビン耐性ヒト癌細胞株におけるdeoxycytidine kinaseの不活化

    齋木由利子, 吉野優樹, 福重真一, 砂村真琴, 石田晶玄, 元井冬彦, 江川新一, 海野倫明, 堀井明

    第７１回日本癌学会学術総会　2012/09/19

  28. miR-34a発現低下は鼻副鼻腔癌におけるシスプラチン化学療法の予後規定因子である

    小川武則, 齋木由利子, 志賀清人, 松浦一登, 加藤健吾, 西條茂, 小林俊光, 福重真一, 堀井明

    第７１回日本癌学会学術総会　2012/09/19

  29. メチルCpG配列を標的とする転写活性化とマイクロアレイの組み合わせはヒトがんにおける低メチル化遺伝子の同定を可能にする

    福重真一, 陳娜, 堀井明

    第７０回日本癌学会学術総会　2011/10/03

  30. 大腸癌における新規癌抑制遺伝子NDRG2のエピジェネティックな発癌制御機構

    山村明寛, 堀井明, 三浦康, 唐沢秀明, 阿部佳子, 内藤剛, 小川仁, 木内誠, 矢崎伸樹, 羽根田祥, 渡辺和宏, 大沼忍, 佐瀬友彦, 鈴木秀幸, 斉木由利子, 福重真一, 柴田

    第74回大腸癌研究会　2011/01/21

  31. ヒトがんにおいて各メチルCpG結合ドメインは異なるメチル化遺伝子を標的とする能力をもつ

    福重真一, 堀井明

    第６９回 日本癌学会学術総会　2010/09/22

  32. 癌抑制遺伝子NDRG2のエピジェネティックサイレンシングは消化器発癌を進行させる

    山村明寛, 三浦康, 阿部佳子, 福重真一, 斉木由利子, 木内誠, 安藤敏典, 唐沢秀明, 金子直征, 佐瀬友彦, 柴田近, 佐々木巌, 堀井明

    第６９回 日本癌学会学術総会　2010/09/22

  33. メチル化で発現抑制された遺伝子を特定する新規の方法「MeTA」の膵癌解析への応用

    清水秀幸, 福重真一, 堀井明

    第99回 日本病理学会総会　2010/04/27

  34. Identification of novel targets for aberrant methylation in pancreatic cancer by a newly developed method “methyl-CpG targeted transcriptional activation (MeTA). International-presentation

    Fukushige, S, Shimizu, H, Sato, Y, Horii, A

    101st American Association for Cancer Research Annual Meeting　2010/04/17

  35. S100A4 regulates tumor growth, motility, and invasion in pancreatic cancer. International-presentation

    Sekine, H, Tabata, T, Tsukamoto, N, Yamamura, A, Abe, K, Yoshino, Y, Sato, K, Chen, N, Umetsu, Y, Sato, D, Gu, Z, Fukushige, S, Shima, K, Motoi, F, Egawa, S, Sunamura, M, Horii, A

    101st American Association for Cancer Research Annual Meeting　2010/04/17

  36. Expression of the N-myc downstream-regulated gene 2 (NDRG2) is frequently suppressed by promoter hypermethylation in human gastrointestinal and pancreatic cancers. International-presentation

    Yamamura, A, Miura, K, Karasawa, H, Abe, K, Mizuguchi, Y, Jin, G, Gu, Z, Fukushige, S, Kaneko, N, Kinouchi, M, Ando, T, Yazaki, N, Tanaka, N, Sase, T, Shibata, C, Sasaki, I, Horii, A

    101st American Association for Cancer Research Annual Meeting　2010/04/17

  37. 癌抑制遺伝子NDRG2のエピジェネティックサイレンシングは消化器発癌を進行させる

    山村明寛, 三浦康, 木内誠, 安藤敏典, 矢崎伸樹, 田中直樹, 唐澤秀明, 佐瀬友彦, 木村俊一, 福重真一, Zhaodi Gu, 嶋健太郎, 柴田近, 堀井明, 佐々木巌

    第110回日本外科学会定期学術集会　2010/04/08

  38. Identification of epigenetically silenced genes by the novel methyl-CpG-targeted transcriptional activation method in human pancreatic cancer. International-presentation

    Hideyuki Shimizu, Yuko Sato, Akira Horii, Shinichi Fukushige

    The 8th AACR/JCA Joint Conference　2010/02/05

  39. Identification of epigenetically silenced genes by methyl-CpG targeted transcriptional activation in pancreatic cancer.

    Shinichi Fukushige, Hideyuki Shimizu, Akira Horii

    第68回 日本癌学会学術総会　2009/10/01

  40. Methyl-CpG targeted transcriptional activation identifies transcriptionally silenced genes by DNA hypermethylation.

    Yuko Sato, Akira Horii, Shinichi Fukushige

    第68回 日本癌学会学術総会　2009/10/01

  41. S100A4 regulates cell growth and motility in pancreatic cancer cells

    Hitoshi Sekine, Nobukazu Tsukamoto, Takahiro Tabata, Yuki Yoshino, Yukiko Umetsu, Zhaodi Gu, Daisuke Sato, Kentaro Shima, Shinichi Fukushige, Fuyuhiko Motoi, Shinichi Egawa, Makoto Sunamura, Akira Horii

    第68回 日本癌学会学術総会　2009/10/01

  42. Promoter hypermethylation and reduced expression of NDRG2 in gastrointestinal cancers

    Akihiro Yamamura, Koh Miura, Makoto Kinouchi, Toshinori Andou, Nobuki Yazaki, Naoki Tanaka, Hideaki Karasawa, Tomohiko Sase, Shinichi Fukushige, Zhaodi Gu, Chikashi Shibata, Iwao Sasaki, Akira Horii

    第68回 日本癌学会学術総会　2009/10/01

  43. Methyl-CpG targeted transcriptional activation in human cancer cells International-presentation

    Shinichi Fukushige

    Japan-Denmark Joint Workshop "Molecular Cancer Research"　2009/01/19

  44. Methyl-CpG targeted p300 recruitment reactivates the expression of epigenetically silenced tumor suppressor genes

    Shinichi Fukushige, Akira Horii

    the 67th Annual Meeting of the Japanese Cancer Association　2008/10/28

  45. がん抑制遺伝子のエピジェネティックサイレンシングはヒストン修飾の改変によって再活性化される

    福重真一, 近藤恵美子, 堀井 明

    第30回日本分子生物学会年会・第80回日本生化学会大会　2007/12/11

  46. Changes in histone modifications reactivate the expression of epigenetically silenced tumor suppressor genes in cancer cells International-presentation

    Shinichi Fukushige, Emiko Kondo, Akira Horii

    The American Society of Human Genetics 57th Annual Meeting　2007/10/23

  47. Changes in histone modifications reactivate the expression of epigenetically silenced tumor suppressor genes

    Shinichi Fukushige, Emiko Kondo, Akira Horii

    第６６回 日本癌学会学術総会　2007/10/03

  48. GSTP1遺伝子の再発現におけるヒストン脱アセチル化酵素阻害の役割

    福重真一, 近藤恵美子, 堀井明

    第65回日本癌学会学術総会　2006/09/28

  49. メチルCpG結合蛋白MBD2、MBD4を介した転写抑制におけるRET finger proteinの役割

    近藤恵美子, Zhaodi Gu, 堀井明, 福重真一

    第65回日本癌学会学術総会　2006/09/28

  50. Whole Genome Amplificationを利用した膵癌の遺伝子診断

    阿部忠義, 砂村眞琴, 江川新一, 福山尚治, 元井冬彦, 福重真一, 堀井明, 海野倫明

    第61回日本消化器外科学会定期学術総会　2006/07/13

  51. PanIN病変、膵癌のWhole Genome Amplificationを用いたゲノム解析

    阿部忠義, 砂村眞琴, 江川新一, 福山尚治, 元井冬彦, 福重真一, 堀井明, 海野倫明

    第37回日本膵臓学会大会　2006/06/29

  52. Ret finger protein mediates MBD2- and MBD4-dependent transcriptional repression. International-presentation

    Fukushige, S, Gu, Z, Kondo, E, Horii, A

    97th American Association for Cancer Research Annual Meeting　2006/04/01

  53. 癌関連遺伝子の高度メチル化プロモーターにおけるMBD4蛋白の役割

    福重 真一, 近藤 恵美子, Zhaodi Gu, 堀井 明

    第６４回 日本癌学会学術総会　2005/09/14

  54. メチルCpG結合蛋白を介した転写抑制におけるRet finger proteinの役割

    Zhaodi Gu, 近藤 恵美子, 堀井 明, 福重 真一

    第６４回 日本癌学会学術総会　2005/09/14

  55. Cre-lox遺伝子挿入系を用いたp53ミスセンス変異の解析

    近藤 恵美子, 堀井 明, 福重 真一

    第６４回 日本癌学会学術総会　2005/09/14

  56. 高度メチル化プロモーター領域へのMBD4蛋白の関与

    福重真一, 近藤恵美子, Zhaodi Gu, 堀井明

    第63回 日本癌学会学術総会　2004/09/29

  57. メチルCpGを介した転写抑制におけるMBD4とその相互作用蛋白RFPの関与

    近藤恵美子, Zhaodi Gu, 堀井明, 福重真一

    第63回 日本癌学会学術総会　2004/09/29

  58. メチルCpG結合蛋白MBD4による転写抑制活性

    福重 真一, 近藤 恵美子, 堀井 明

    第２６回 日本分子生物学会年会　2003/12/10

  59. メチルCpG結合蛋白MBD4の転写抑制活性

    福重 真一, 近藤 恵美子, 堀井 明

    第６２回 日本癌学会総会　2003/09/25

  60. hMLH1のN末に相互作用する蛋白のスクリーニング

    近藤 恵美子, 堀井 明, 福重 真一

    第６２回 日本癌学会総会　2003/09/25

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Research Projects 20

  1. Elucidation of ZNF750-TWIST1 pathway in prostate cancer metastasis

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2024/04/01 - 2027/03/31

  2. Development of liquid biopsy technology targeting DNA methylation for prostate cancer diagnosis

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2021/04/01 - 2024/03/31

  3. Development of diagnostic technique of pancreatic cancer by using liquid biopsy targeted DNA methylation markers

    Gu Zhaodi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2019/04/01 - 2022/03/31

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    In this study, we examined the development of a liquid biopsy technique targeting DNA methylation of six genes that are hypermethylated and transcriptionally repressed in pancreatic cancer by MeTA, our proprietary hypermethylated gene search method. The amount of cfDNA obtained from 2 mL of plasma from patients with pancreatic diseases including pancreatic cancer averaged 17.4 ng, and higher molecular weight band ladders were not detected. Although DNA methylation is difficult to detect at this time, methods of sampling plasma and methylated DNA enrichment need to be investigated. On the other hand, analysis of LHX6 gene identified by MeTA revealed that transcriptional repression by hypermethylation plays an important role in pancreatic cancer cell growth.

  4. Role of PYCARD, an apoptosis-inducing gene, in prostatic tumorigenesis

    Fukushige Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2018/04/01 - 2022/03/31

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    Using mouse model of prostate cancer, we aimed to clarify the role of PYCARD gene, which shows promoter hypermethylation and inactivation in human prostate cancer. Although further analysis is needed to draw conclusions due to the small number of individuals, the genotype of Pycard may influence precancerous lesions and tumor size in prostate cancer in prostate-specific Pten knockout mice. On the other hand, analysis of human prostate cancer tissue suggested that inactivation of PYCARD may already occur in precancerous lesions and that aberrant DNA methylation of PYCARD is a distinctive feature of prostate cancers with Gleason score >= 7.

  5. Identification of hypermethylated genes associated with increased cell proliferation of prostate cancer using DNA demethyation techniques

    Fukushige Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2015/04/01 - 2018/03/31

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    DNA methylation is associated with the inappropriate transcriptional silencing of cancer-related genes and affects the hallmarks of cancer such as sustaining proliferative signaling, resisting cell death, and activating invasion and metastasis. We found that genome-wide reactivation of hypermethylated genes suppressed growth of prostate cancer cell line LNCaP and induced apoptosis. We searched for apoptosis-inducing genes whose promoters were hypermethylated in prostate cancers. As a result, we found that the promoter region of PYCARD, one of apoptosis-inducing factor, were highly methylated in a cancer-specific manner (46/51: 90%). In addition, PYCARD expression induced apoptosis in LNCaP cells. These results suggest that aberrant promoter hypermethylation of PYCARD plays an important role in prostatic tumorigenesis.

  6. Development of DNA demethylation technology in cancer cells and its application for discovering biomarkers for early diagnosis

    FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2012/04/01 - 2015/03/31

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    We have developed a novel method that globally demethylates and reactivates hypermethylated genes by using a fusion gene comprising of methyl-CpG binding domain (MBD) and the catalytic domain of Ten-eleven translocation protein 1 (TET1-CD). This method is useful for identifying hypermethylation-mediated silenced genes in cancer cells. We have applied it to prostate and colorectal cancer cell lines and found that DNA demethylation suppressed the growth of cancer cells. These results suggest that DNA methylation confers the ability of cancer cells to facilitate the growth and DNA methylation of gene(s) responsible for cell growth may be useful as a biomarker.

  7. The search for DNA methylation-based biomarkers in cancer

    FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2009 - 2011

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    We have developed a novel method called"microarray coupled with methyl-CpG targeted transcriptional activation" (MeTA-array for short), which is useful in identifying hypermethylation-mediated silenced genes in cancer. The majority of genes identified by this method had more stringent criteria of CpG islands at the promoter region at high frequency. We have applied MeTA-array to pancreatic cancer cells and identified several genes hypermethylated in primary pancreatic cancers in a tumor-specific manner. These genes should be useful as candidates for tumor-specific biomarkers.

  8. Functional analysis of MBD proteins in epigenetic silencing of cancer-related genes

    FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (B)

    Institution: Tohoku University

    2006 - 2007

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    Epigenetic modifications such as DNA methylation and histone modification play crucial roles in the pathogeneses of cancer by transcriptional silencing of tumor suppressor genes. To clarify the mechanism(s) in which methyl-CpG binding domain (MBD) proteins repress transcription, we analyzed proteins interacting with MBD4 and developed the methods to search cancer-related genes regulated by MBD proteins. First, we found that Ret finger protein (RFP) interacts with MBD4 and enhances MBD2-and MBD4-dependent transcriptional repression. Because RFP has been detected at high levels in a variety of tumor cell lines as well as testis, and embryos, RFP may have an important role in the enhancement of transcriptional repression through MBD proteins in tumorigenesis, spermatogenesis, and embryogenesis. We next found that NF_KB transcriptional activation domain linked to MBD (NF_κB(AD)-MBD) reactivated the expression of densely methylated MLH1 genes in HEK293T and AN3CA cells without demethylation. This phenomenon seems to be general since we observed the transcriptional reactivation in all ten cancer-related genes analyzed so far. Furthermore, microarray analyses using HEK293T cells transfected with HF_κB(AD)-MBD showed re-expression of a variety of cancer-related genes such as CDH1. Therefore, this approach may provide an efficient method to screen epigenetically silenced cancer-related genes mediated by MBD proteins.

  9. 安定なRNA干渉を可能にするステーブル細胞株作製技術の開発

    福重 真一

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 萌芽研究

    Institution: 東北大学

    2005 - 2006

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    本研究ではCre-lox遺伝子挿入系を用い、ゲノム上の特定部位(前もって導入したlox部位)に二本鎖のsiRNAを発現するコンストラクトを正確に単一コピー挿入することによって、確実にRNA干渉をおこなう細胞株の作製を目的とした。 本年度は、Ltk-細胞に単一コピーのlox位を含む細胞株、73-14を用い、マウスRet finger protein (RFP)に対するRNA干渉コンストラクトをCre-lox遺伝子挿入系により導入したステーブル細胞株を作製し、その効果を解析した。まず、73-14にRFPのRNA干渉コンストラクトを含むloxターゲティングベクターとCre発現プラスミドをトランスフェクションし、G418での薬剤選択により14個の組換え体を得た。これらの組換え体からゲノムDNAを抽出し、種々のPCRで組換え部位、RNA干渉コンストラクトの存在を確認した。次に、ネオマイシン耐性遺伝子をプローブとしてサザンハイブリダイゼーションをおこない、14クローンすべてがゲノム中のlox部位に正確にRFPのRNA干渉コンストラクトをもつことを確認した。さらに、ターゲティングベクター中のCMVプロモーターをプローブとしサザンハイブリダイゼーションをおこない、14クローン中、11クローンが単一コピーの組換え体であり、残り3クローンは2コピー以上の組換え体であることが判明した。 ウエスタンブロティング法により14クローンのRFP干渉による蛋白レベルでの変化を解析した結果、どのクローンでも親株に比べ約50%のノックダウンが観察された。興味深いことにRNA干渉コンストラクトを2コピー以上もつ3個の組換え体でも単一コピーをもつ残り11個の組換え体とノックダウン率に大きな違いが見出せなかった。このことは、コピー数よりもRNA干渉を引き起こすshRNAの塩基配列を変える方がより効果的にRNA干渉をおこなえる可能性を示すと考えられる。

  10. Mechanism of transcriptional silencing mediated by the methyl-CpG binding protein MBD4 and RET finger protein

    KONDO Emiko, FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2005 - 2006

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    Aberrant gene silencing in tumors via methylation of CpG islands in the promoter of many cancer-related genes affects virtually every step in tumor progression. In this process, the methyl-CpG binding domain (MBD) proteins play an essential role in transmitting epigenetic information to downstream regulatory proteins. Among the five MBD proteins identified so far, MBD4 has been the only exception; it has long been thought to be a DNA repair protein. In this study, we demonstrated that MBD4 also has the ability to repress transcription through methyl-CpG sequences. MBD4 is involved in HDAC-dependent repression and directly binds to HDAC1 and corepressor SIN3A. Furthermore MBD4 specifically binds to highly methylated promoters of CDKN2A and MLH1 genes. In order to further investigate the role of MBD4 in methylation-based transcriptional repression, a yeast two-hybrid screening was performed, and the RET finger protein (RFP) was found to be one of the major proteins that interact with the transcriptional repression domain in MBD4. The effect of the MBD4-mediated transcriptional repression in methylated CDKIV2A and MLH1 promoters was extremely enhanced by the overexpression of RFP. Because RFP has been detected at high levels in a variety of tumor cell lines as well as testis, and embryos, RFP may have an important role in the enhancement of transcriptional repression through MBD proteins in tumorigenesis, spermatogenesis, and embryogenesis.

  11. Functional analysis of DNA mismatch protein hMLH1 using germline mutants

    FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    2001 - 2002

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    Hereditary nonpolyposis cotorectal cancer (HNPCC) is the most common form of familial cotorectal cancer and is associated with germline mutations in one of the DNA mismatch repair genes. Among these genes, the hMLH1 gene is the most frequently damaged, and high frequency of the missense mutation (about onethird of all the mutations) is a particularly characteristic feature. Therefore, it is important to clarify whether or not these missense mutations are in fact pathogenic. In this study, we have developed an assay to distinguish between pathogenic mutations and polymorphisms inb hMLH1 variants. We first surveyed the database (http://www.nfdht.nl) presented by the International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer (ICG-HNPCC) and chose 23 missense variants of the hMLH1 protein. We used the yeast two-hybrid assay with hPMS2 or hEX01 as bait proteins to measure the function of the protein and found that 18 (78%) of 23 variants showed significant decreased β-gal activities in comparison with the wild type hMLH1. Two of the remaining five variants were thought to be rare polymorphisms. Hence the yeast two-hybrid assay is an extremely simple and useful tool for evaluating the biological significance of the variant hMLH1 proteins. We further analyzed 14 truncating mutations (11 frameshtft and 3 nonsense mutations) and one in-frame 3-bp deletion; these mutants were chosen because these still remain some or most of the Cterminai portion for hPMS2-interacting domain. AH of these 15 mutants also showed extremely low levels of β-gal activities. Our present results suggest that these dysfunctions are due to either lack in the interacting domain with associating proteins at the C-terminus or con formationai changes caused mainly by the N-terminal abnormalities. In conclusion, the yeast two-hybrid assay is a simple and reliable system for accurate diagnosis of the hMLH1 alterations.

  12. Reseearch on Genetic Alterations in the Development and Progression of Human Pancreatic Cancer

    HORII Akira, HOSHI Masato, FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (B)

    Institution: Tohoku University

    2000 - 2001

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    Six frequently lost regions in pancreatic cancer were analyzed for association between poor prognosis and LOH ; in three regions (12q, 17p, and 18q) significant association. Among these, 18q harbors SMAD4, one of the key genes in pancreatic carcinogenesis. IPMT is recognized as one of the premalignant lesions of pancreatic ductal cells, and frequent loss of 18q at the SMAD4 locus has been observed. However, SMAD4 did not alter and its protein product expressed. Introduction of SMAD4 did not affect cell growth in vitro, irrespective of SMAD4 status. On the other hand, introduction of chromosome 18 significantly suppressed cell growth in vitro in both cells with and without inactivation of SMAD4. These observations strongly suggest the existence of novel tumor suppressor gene, near but distinct from SMAD4, that plays an important role in the early stage of pancreatic tumorigenesis. On 12q, we found the DUSP6 gene, the protein product of which works as an ERK specific phosphatase. Expression of this gene was frequently suppressed in pancreatic cancer, and adenovirus mediated introduction caused apoptosis in pancreatic cancer cells. BAI1 is one of the important protein that upregulated by p53. It works as angiogenesis inhibitor and expresses specifically in brain. We introduced BAI1 in pancreatic cancer cells using adenovirus vector and observed growth suppression in vivo by means of suppressed angiogenesis; there is a possibility of gene therapy utilizing tumor dormancy. 1p36-p35 is one of the hot spots for LOH in a variety of human cancers. We constructed a 35-Mb BAC-based contig in this region. The RIZ gene encoding the RB interacting zinc finger protein was in this region and mutation analysis revealed the frequent frameshift mutation at the microsatellite in the coding region of the gene in MSI-H tumors of the colorectum, stomach, and endometrium as well as pancreas. RIZ may be one of the candidates for mutation in MSI-H pancreatic cancers.

  13. Identification of tumor suppressor genes by means of a genomic clone-transfer

    FURUKAWA Toru, SUNAMURA Makoto, FUKUSHIGE Shinichi, HORII Akira, SHIBUYA Kazchiko

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (C)

    Institution: Tohoku University

    1999 - 2000

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    The pancreatic cancer cells are in high frequency of loss of heterozygosity in some chromosomal allels including 1p, 6q, 9p, 12q, 17P, and, 18q. In these deleted alllels, tumor suppressor genes may exist. Aim of this study was to identify tumor suppressor genes in the deleted regions in the pancreatic cancer by means of a genomic clone-transfer. We focused on 6q, 12q and 18q because no responsible genes have been identified yet despite high frequency of LOH.To find genomic clones, human yeast and bacterial artificial chromosome (YAC, BAC) libraries were screened. Identified clones were constructed in contig to cover the deleted regions. For the deleted region at 6q21, a complete BAC contig to span 1-Mb was constructed. We identified 42 ESTs in the region. The contig was available at http : //www-alis.tokyo.jst.go.jp/HGS/plmap.pl? PlID=14.For 12q21, a contig comprised of 21 BAC clones which could span 3-Mb was constructed. Forty novel STSs and 10 ESTs were identified. We identified several novel genes in the deleted regions including hGnT-IV-H, hHDC, and TUB1. Information and references for these genes were available in Genbank/EMBL/DDBJ.To know genetic status of pancreatic cancer cells to be transferred, we analyzed cancer-related genes including KRAS, p16, p53 and SMAD4. To know a status of 18q, we analyzed and identified high frequency of LOH of 18q in intraductal papillary mucinous tumors, which was thought to be an early lesion of conventional pancreatic cancer, with low frequency of mutation of SMAD4. Since loss of 18q was found to span broad area, transfer of chromosome 18 was chosen and carried out to compensate its function in the pancreatic cancer cells. The transferred clone revealed limited growth in vitro and tumorigenesity in vivo.

  14. Genetic alterations involved in initiation and progression of human cancer

    HORII Akira

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research on Priority Areas

    Institution: Tohoku University School of Medicine

    1999 - 1999

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    1. Among regions of frequent chromosomal aberrations, loss of three chromosomal regions, 12q, 17p, and 18q, associated with poor prognosis in human pancreatic cancer. Ade noviral mediated delivery of the SMAD4 gene in pancreatic cancer cell lines with homozygous deletion of SMAD4 did not show any suppression of cell growth. We previously reported that loss of 18q is an early event in pancreatic carcinogenesis. There is a possibility that mutation of the SMAD4 gene is responsible for the initial step of pancreatic carcinogenesis as well as prognosis defining factor. However there is a possibility of unknown tumor suppressor gene on 18q that is distinct from SMAD4. 2. We and others previously reported frequent somatic mutation of the PTEN gene in endometrial cancer. We attempted a trial of gene therapy by adenovirus mediated introduction of this gene in endometrial cancer cell lines with two-hit mutation of this gene. The trial was successful in vitro, and apoptosis was induced in tumor cells after introduction of normat copy of PTEN.However, the attempt was not successful in vivo. These results suggested that the PTEN gene is a good candidate for gene therapy in human endometrial cancer after appropriate improvement. 3. In human lung cancer, frequent loss of 10q at the DMBT1 locus was found. Moreover, mutation as well as suppression of expression was found in this gene. There is a possibility that DMBT1 acts as the tumor suppressor gene in human lung carcinogenesis. 4. In neuroblastoma, allelic loss was studied in 14q and identified a 500-kb region of common deletion on 14q32. A BAC contig harboring the deleted region was also constructed. On the other hand, a break point on 1p32 that occurred in a neuroblastoma patient with constitutional reciprocal translocation was also cloned. We are attempting to isolate the genes responsible for neuroblastoma.

  15. Establishment of novel efficient methods for pancreatic cancer patients based on genetic abnormality.

    MATSUNO Seiki, HORII Akira, SUNAMURA Makoto, FUKUSHIGE Shinichi, FURUKAWA Toru, SHIBUYA Kazuhiko

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (A)

    Institution: Tohoku Uniersity

    1998 - 1999

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    1.Among regions of frequent chromosomal aberrations, loss of three chromosomal regions, 12q, 17p, and 18q, associated with poor prognosis in human pancreatic cancer. 2.Adenoviral mediated delivery of the SMAD4 gene in pancreatic cancer cell lines with homozygous deletion of SMAD4 did not show any suppression of cell growth. We previously reported that loss of 18q is an early event in pancreatic carcinogenesis. There is a possibility that mutation of the SMAD4 gene is responsible for the initial step of pancreatic carcinogenesis as well as prognosis defining factor. However, there is a possibility of unknown tumor suppressor gene on 18q that is distinct from SMAD4. If this is the case, then the SMAD4 gene associate with poor prognosis that disrupt at the late stage, and disruption of the unknown putative tumor suppressor gene plays at the early stage tumors. 3.In 6q and 12q, we identified three and two commonly deleted region. We further narrowed down the regions, and constructed BAC contigs in three regions; one on 6q and two on 12q. We isolated and characterized the DUSP6 and TDG genes on 12q and found that expressions of both genes were significantly reduced. We are now further characterizing the gene to investigate their roles in pancreatic carcinogenesis. 4.We demonstrated inhibition of angiogenesis by MMP inhibitors and IL-12. These results will lead to establishment of clinical application through regulation of tumor angiogenesis. We also established mutant adenovirus that only can survive in cells with disrupted p53. This virus is a good candidate for gene therapy of pancreatic cancer.

  16. DNA除去修復機構とその修復欠損の分子病態

    田中 亀代次, 関 周司, 安井 明, 花岡 文雄, 福重 真一, 山泉 克

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 特定領域研究(A)

    Institution: 大阪大学

    1996 - 1999

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    田中は、A群色素性乾皮症蛋白質、A及びB群コケイン症候群蛋白質、さらには、RNA polymerase IIと結合し、「転写と共役したヌクレオチド除去修復」及び転写機構に関与するXAB2蛋白質を同定し、XAB2が含まれる蛋白質複合体を精製した。質量分析法により、その構成因子をコードする遺伝子を同定した。マウスXAB2遺伝子をクローニングしてのち、XAB2遺伝子のノックアウトは胎児致死をもたらすことを明らかにした。RLGS法により、c-mycによるアポトーシス抑制に関わる転写因子がXPA欠損マウスの皮膚癌細胞で高発現していることを見つけた。花岡は、XP-V群の損傷DNA複製欠損を相補する蛋白質をHeLa細胞核抽出液より精製し、シクロブタン型ピリミジン2量体を乗り越えて正しく複製できる新規のDNAポリメラーゼであることを明らかにした。5例のXP-V群患者細胞において変異が確認されたことから、これがXPV責任遺伝子であることを証明した。安井は、8-オキソグアニンと誤対合したアデニンを切り出すヒトhMYH蛋白質が、ミトコンドリアと核に存在することを見つけた。また、光回復酵素のマウスホモログが日周リズムに必須の蛋白であることを示した。関は、大腸菌endonuclease IIIの哺乳類ホモログNTHL1遺伝子と結節性硬化症遺伝子TSC2とは、head to headで近接して存在し、両遺伝子の転写は基本的には両方向性のコアプロモーターで調節されていることを明らかにした。山泉は、電離放射線照射後のp53蛋白質セリン15のリン酸化がその安定化・活性化に重要であることを明らかにした。また、DDB2遺伝子に変異を持つXP-E群患者を見出し、この患者細胞ではピリミジン2量体の修復は正常だが6-4光産物の修復に遅れが認められることから、DDB2が6-4光産物の後期の修復に関与している可能性を示唆した。福重は、遺伝性非腺腫症性大腸癌の家系に見られる59種のhMLH1胚細胞変異のうち、50種でhMLH1-hPMS2相互作用の強い阻害を観察し、これがミスマッチ修復機構破壊の主な原因と考えた。

  17. 環状酵母人工染色体(YAC)移入による癌抑制遺伝子の同定

    福重 真一

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 萌芽的研究

    Institution: 東北大学

    1997 - 1998

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    本研究は、遺伝子機能の欠損を原因とする疾患において、その原因遺伝子単離へのアプローチとして、欠損遺伝子を含むと考えられる環状のYACDNA断片を作製後、細胞に導入し、病態が修正されるかどうか検討することにある。本年度は、まず、マイクロサテライトマーカーを用いたアレロタイプ解析により膵癌の癌抑制遺伝子の候補領域として、第6番染色体長腕における共通欠失領域を3箇所同定し、その中でも最も高頻度(69%)のLOHを認めた領域(D6S449-D6S283)を6q21の500-kbに絞り込むことに成功した。また、胃癌、肺癌においてもアレロタイプ解析により16q24に共通欠失領域を見出した。次に、昨年まで検討していたCre-lox部位特異的組換えにより、環状のYACクローンを得るという方法から、より直接的で簡便な方法としてTransformation Associated Recombination(TAR)法によりヒトのゲノムDNAからマイクロサテライトマーカー周辺の塩基配列の情報だけを頼りに共通欠失領域を含む環状YACクローンを得ることを考案した。TAR法ではYACベクターに共通欠失領域を決める2つのマーカー周辺の塩基配列を挿入し、ヒトゲノムDNAと共に酵母細胞に導入し、酵母の高率な相同組換えを利用して2つのマーカー間のクローニングを行なう。しかしながら、今現在、期待されるYACクローンを得るに至っていない。その原因として、マイクロサテライトマーカー周辺の知られている塩基配列が200-300bpと短いため、酵母において特異的な領域での相同組換えの頻度が低いことが考えられる。マーカー周辺の塩基配列を決定し、より長いユニークなDNA断片をベクターに挿入するなどの改良を加えることが今後の課題である。

  18. Identification of tumor suppressor genes in human pancreatic cancer

    HORII Akira, FUKUSHIGE Shinichi, SUNAMURA Makoto

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Scientific Research (B)

    Institution: Tohoku University

    1995 - 1996

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    We first searched for imbalances of copy number in human pancreatic cancer cells using more than 100 primary tumor and corresponding normal tissues as well as 12 cell lines by (i) LOH study with microsatellite markers of all chromosome arms except for short arms of acrocentric chromosomes and sex chromosomes and (ii) comparative genomic hybridization (CGH). Frequent gains of chromosome arms 8q and 20q, and losses of 1p, 6q, 9p, 12q, 17p, and 18q were observed. We further focused on allelic loss of chromosome arm 12q and constructed a detailed deletion map. Finally we identified a 1-cM region of common allelic loss. We further constructed a contig in this region with YAC and BAC clones, and found that a YAC colne of 790 kb in size harbored the whole region of common allelic loss. We are now constructing a cosmid contig in this region. Region of the gain of chromosome 8q overlapped with the region of the c-myc oncogene. Region of the gain of chromosome 20q was evaluated for its precise region and copy number by fluorescence in situ hybridization (FISH). Copy number was increased by two-fold, and the region was overlapped with that of breast cancer. Involvement of DNA mismatch repair error was also analyzed in this study. Frequent microsatellite instability (MI) was observed (13% of the tumors). None of the mutations in hMSH2 and hMLH1 were observed in tumors with MI+.Moreover, somatic mutations of neither the transforming growht factor (TGF) beta receptor type II (RII) gene nor the insulin-like growht factor II receptor (IGFIIR) gene were found which were frequently observed in colorectal, gastric, and endometrial cancers with MI+. These results suggested that mutations of these genes do not play an important role, if any, in pancreatic carcinogenesis.

  19. ヒト染色体の分離と細胞への導入

    福重 真一

    Offer Organization: 日本学術振興会

    System: 科学研究費助成事業

    Category: 奨励研究(A)

    Institution: 大阪大学

    1987 - 1988

  20. Development of chromosome sorting techniques using a dual laser chromosome sorter

    MATSUBARA Kenichi, OCHIYA Takahiro, FUKUSHIGE Shinichi

    Offer Organization: Japan Society for the Promotion of Science

    System: Grants-in-Aid for Scientific Research

    Category: Grant-in-Aid for Developmental Scientific Research

    Institution: Osaka University

    1986 - 1988

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    We have developed chromosome sorting techniques using a dual laser chromosome sorter to analyze the structure of human chromosomes. Human chromosomes stained with Hoechst 33258 (Ho) and chromomycin A3 (CA3) flowed sequentially through two laser beams, one adjusted to 457.9nm to excite CA3 and the other adjusted to 351.1-363.8nm to excite Ho. The Ho and CA3 contents of individual chromosomes were determined by measuring the intensities of fluorescence that were emitted as they passed two laser beams. These measurements were accumulated to form fluorescence intensity distributions. Sorting techniques have evolved from the ability to sort 10-20 human chromosome fractions with a single laser system to sorting 21 fractions including 15 unique types with a dual laser system. dFurthermore, chromosome types that cannot be sorted at high purity from chromosomes isolated from human cells can be sorted from human-hamster hybrid cell lines containing one or a few human chromosomes.

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  1. がんにおけるDNAメチル化異常へのMBD蛋白の関与

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    DNAメチル化は遺伝子の転写抑制に密接に関連するエピジェネティック変化であり、がんではがん抑制遺伝子の高メチル化、がん遺伝子の低メチル化ががんの発生・進展に大きな影響を及ぼす。遺伝子転写抑制の主要なメカニズムとしてメチルCpG結合（MBD）蛋白の遺伝子メチル化プロモーターへの結合、ヒストン脱アセチル化酵素などヒストン修飾蛋白のリクルートが重要であると考えられる。MBD蛋白にはMBD1〜4、MeCP2の計５種類が存在するが、それぞれの塩基配列特異性や遺伝子特異性についてはまだ十分な情報がない。本研究では、我々が開発したMeTA-array（microarray coupled with methyl-CpG targeted transcriptional activation）という方法を駆使し、がん細胞においてそれぞれのMBD蛋白により転写抑制されるメチル化候補遺伝子を同定し、臨床検体におけるメチル化状況を検討する。すでに12種の膵がん細胞株とコントロールとして正常膵管上皮細胞株にMBD2のMBDを用いたMeTA-arrayを適用し、半数の膵がん細胞株に共通に見られる高メチル化候補遺伝子31種と低メチル化候補遺伝子19種を同定した。数種の高メチル化候補遺伝子の解析から臨床検体でも、高メチル化が高頻度で見られることが判明し、MeTA-arrayの有用性が示された。これらの解析を他のMBDを用いたMeTA-arrayでも同様に進めて行く。また、MBD蛋白のノックダウン等の手法を用い、各メチル化遺伝子の転写抑制にこれらがどのような役割を果たしているのか明らかにしていく。

  2. がん早期診断へ向けたメチル化miRNA遺伝子による遺伝子診断セットの開発

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    【目的】がんの診断マーカーとなるメチル化miRNA遺伝子の効率的な探索法の開発と診断への応用を目的とする。【内容】膵がんを例として、メチルCpG配列を標的とする転写活性化（MeTA）とmiRNAのマイクロアレイを組み合わせ、３種の膵がん細胞株でメチル化miRNA遺伝子が得られる効率を調べる。また、すべての細胞で共通に発現異常を示すメチル化miRNA遺伝子を見つけ、多段階発がん過程におけるメチル化時期を決定し、診断への応用の可能性を検討する。