2000 Fiscal Year Final Research Report Summary
Elucidation of activation mechanism of MN/CA9 in human renal cell carcinomas
| Project/Area Number |
11670224
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | NARA MEDICAL UNIVERSITY |
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Principal Investigator |
CHO Masaki Dept.Urol.Nara Med.Univ. Research Associate, 医学部, 助手 (90285362)
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| Co-Investigator(Kenkyū-buntansha) |
KONISHI Noboru 2nd Dept.Pathol.Nara Med.Univ. Professor, 医学部, 教授 (20145832)
HIRAO Yoshihiko Dept.Urol.Nara Med.Univ. Professor, 医学部, 教授 (00133207)
UEMURA Hirotsugu Dept.Urol.Nara Med.Univ. Assistant Professor, 医学部, 講師 (90213397)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Renal cell carcinoma / MN / CA9 / methylation / DNAメチル化 |
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| Research Abstract |
Correlation between MN/CA9 expression and hypomethylation of MN/CA9 5'flanking region in human renal cell carcinomas
Hypomethylation of the CpG at -74 bp of MN/CA9 was strongly correlated with MN/CA9 expression in both human renal cell carcinoma (RCC) cell lines and human RCC tissues. Methylation status was examined by bisulfite genomic sequencing protocol. MN/CA9 expression was examined by RT-PCR.
Induction of MN/CA9 expression by treatment with a demethylating agent
Treatment with the demethylating agent 5-aza-2'-deoxycytidine resulted in activation of the MN/CA9 gene in MN/CA9 negative RCC cell lines.
Identification of MN/CA9 promoter region
DNAs of MN/CA9 5' flanking region (-1246 / +42 bp) were ligated into luciferase reporter plasmids. Deletion analyses revealed that 5' end of minimal MN/CA9 promoter was located between -158 bp and -58 bp.
Effect of in vitro DNA methylation on promoter activity
MN/CA9 promoter (-158 / +42 bp) / luciferase reporter plasmids were treated with CpG methylases (SssI or HhaI) and transfected into MN/CA9 positive RCC cell lines. All CpGs of MN/CA9 promoter were methylated by SssI, and only CpG at -74 bp was specifically methylated by HhaI.Activity of MN/CA9 promoter was strongly suppressed by treatment with both SssI and HhaI.
These data suggest that hypomethylation of the CpG at -74bp in MN/CA9 promoter may have a major role in MN/CA9 expression in human renal cell carcinomas.
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[Publications] Cho, M., Kitahori, Y., Hiasa, Y., Nakagawa, Y., Uemura, H., Hirao, Y., Ohnishi, T., Yoshikawa K.and Oosterwijk, E.: "Activation of the MN/CA9 gene is associated with hypomethylation in human renal cell carcinoma cell lines."Molecular Carcinogenesis. 27. 184-189 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Uemura, H., Yoshikawa, M., Shimizu, K., Cho, M., Nakagawa, Y., Ozono, S., Hirao, Y., Yoshikawa, K.: "MN/CA IX as a potential target for active specific immunotherapy in renal cell carcinomas."Biotherapy. 14. 487-489 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Uemura, H., Nakagawa, Y., Hirao, Y., Okajima, E., Yoshikawa, K., Oosterwijk, E.: "Expression of MN antigen in urologic cancer : potential tool as a therapeutic target."Aktuel Urology. 31. 52-54 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Uemura, H., Cho, M., Nakagawa, Y., Shimizu, K., Yoshikawa, M., Kim, S., Hirao, Y.: "MN/CA IX antigen as a potential target for renalcell carcinoma."Hinyoukika Kiyou. 46. 745-748 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Uemura, H., Nakagawa, Y., Yoshida, K., Saga, S., Yoshikawa, K., Hirao, Y.and Oosterwijk, E.: "MN/CA IX/G250 as a potential target for immunotherapy of renal cell carcinoma."British J.Cancer. 81. 741-746 (1999)
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「研究成果報告書概要(欧文)」より
