Analysis of the gene expression for multidrug-resistance-associated protein in urogenital cancers by DNA microarrays

DNA微阵列分析泌尿生殖系统肿瘤多药耐药相关蛋白基因表达

• 批准号: 12671526
• 负责人: EHARA Hidetoshi
• 金额: $ 2.05万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671526/
• 关键词: urogenital cancers; anticancer-resistance; DNA chip; real-time RT-PCR; chemosensitivity test

To develop the new DNA microarrays superior to chemosensitivity test in urogenital cancers, we conducted a series of experiments on cell lines. Before the design of DNA chips, we synthesized a series of paired primers against genes for anticancer drug resistance associated proteins. We designed these primers to use real-time RT-PCR, simultaneously. After the fundamental experiments, we measured the expression of MDR1 mRNA by a quantitative RT-PCR (LightCycler, Roche Co.) in cancerous samples of patients undergoing surgery or biopsy (n=504), and compared with chemosensitivity test (MTT assay) in clinical specimens of uroepithelial carcinomas and renal cell carcinomas (n=43). Despite this high incidence of MDR1 expression in renal cell carcinoma no obvious association with clinicopathological parameters was found. Further, we did not find the significant associations between MOR1 expression and clinicopathological findings in prostate cancers and uroepithelial cancers. The measurement of MDR1 expression of uroepithelial cancers did not associated with the degree of resistance to anticancer drugs including doxorubicin.

为了研制一种新的、优于泌尿生殖道肿瘤化疗药物敏感性检测的上级DNA微阵列，我们进行了一系列的细胞系实验。在设计DNA芯片之前，我们合成了一系列针对抗癌药物耐药相关蛋白基因的成对引物。我们设计了这些引物，同时使用实时RT-PCR。基础实验结束后，我们通过定量RT-PCR（LightCycler，Roche Co.）测量了MDR 1 mRNA的表达。对504例手术或活检患者的癌组织进行了MTT法检测，并与43例尿路上皮癌和肾细胞癌的临床标本进行了药敏试验（MTT法）比较。尽管肾细胞癌中MDR 1的表达率很高，但与临床病理参数无明显相关性，此外，我们在前列腺癌和尿路上皮癌中未发现MOR 1表达与临床病理结果的显著相关性。测量尿路上皮癌的MDR 1表达与抗癌药物（包括阿霉素）的耐药程度无关。