| Project/Area Number |
11670571
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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 |
Respiratory organ internal medicine
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
INASE Naohiko School of Medicine, Tokyo Medical and Dental University, Lecturer, 医学部・附属病院, 助手 (60262185)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | malignant mesothelioma / gene therapy / tumor-specific expression / specific promoter / keratin 19 / calretinin / suicide gene / keratin19 / calretinin / 腫瘍特異的プロモーター / 悪性胸膜中皮種 |
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| Research Abstract |
To achieve the specific expression of a transfected suicide gene in malignant mesothelioma cells, we applied the enhancer-promoter fusion sequence of the keratin 19(K19)gene. Northern blot analysis of three mesothelioma cell lines demonstrated that K19 mRNA was expressed most abundantly in the H2052 mesothelioma cell line. Subsequently, in a luciferase reporting assay, K19 promoter(260 bp)exhibited higher promoter activity in H2052 cells than in the other two cell lines. In addition, ligation of a 3' enhancer(80 bp)of the K19 gene to upstream of the K19 promoter sufficiently enhanced the promoter activity. After transfecting an expression vector containing the K19 enhancer-promoter bound thymidine kinase gene(pK19-TK)into the H2052 cells, the pK19-TK transfected cells become more sensitive to GCV than non-transfected cells in vitro and in vivo. The K19 enhancer-promoter sequence seemed to be specific and efficient enough for the expression of the transfected suicide gene in malignant mesothelioma cells.
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