| Project/Area Number |
15591692
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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 |
Urology
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| Research Institution | Kochi University |
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Principal Investigator |
INOUE Keiji Kochi Medical School, Associate Professor, 医学部, 助教授 (00294827)
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| Co-Investigator(Kenkyū-buntansha) |
KARASHIMA Takashi Kochi Medical School, Research Associate, 医学部, 助手 (60304672)
SHUIN Taro Kochi Medical School, Professor, 医学部, 教授 (80179019)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | DNA methylation / metastasis-related gene / transitional cell carcinoma / 移行上皮癌 / E-cadherin / 膀胱移行上皮癌 / 浸潤 / 転移 |
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| Research Abstract |
Purpose : To assess the role of DNA methylation of metastasis-related genes (matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinase (TIMP) and E-cadherin) promoter region in transitional cell carcinoma (TCC).
Experimental Design : The following 9 human TCC cell lines were used :T24, EJ-1,HT-1197,RT4,253J-P,253J-BV, UM-UC-14,NS and KMBC-2. Normal renal proximal tubule cell line RPTEC and normal human umbilical vein endothelial cell line HUVEC was cultured in the recommended growth medium. 22 TCCs and matched adjacent normal tissues were obtained from 22 Japanese TCC patients by surgical resection at the Kochi Medical School. The DNA methylation of metastasis-related genes promoter region in these TCCs was identified by methylation specific PCR (MSP) method (E-cadherin) and combined bisulfite restriction analysis (COBRA) method (MMP2,MMP9,TIMP1,TIMP2). The induction of gene re-expression was also examined by treatment with 5-aza-deoxycytidine (5-aza) and trichostatin A (TSA).
Results : E-cadherin RNA expression was not detected by RT-PCR in T24 and EJ-1, and also TIMP2 in HT-1197 and KMBC-2. Each gene DNA methylation was detected in these cell lines. There was no difference of metastasis-related genes DNA methylation in other cell lines and surgical specimens. E-cadherin gene re-expression was induced by treatment with 5-aza-deoxycytidine (5-aza) and/or trichostatin A (TSA) in T24 and EJ-1, but TIMP2 gene was not in HT1197 and KMBC-2.
Conclusions : DNA methylation of the promoter region may be involved in the regulation of E-cadherin RNA expression. However, the role of DNA methylation of the promoter region in other metastasis-related genes (MMP and TIMP) could not be clarified in present study.
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