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目的:以大规模平行测序技术(massively parallel signature sequencing,MPSS)筛选人肾细胞癌(renal cell carcinoma,RCC)和癌旁组织中差异表达的microRNAs(miRNAs),并验证差异表达miRNAs之一的miR-660在人RCC中的表达。方法:MPSS检测10例RCC组织及相应癌旁组织中miRNAs的表达,筛选RCC组织中差异表达miRNAs。RT-PCR检测5例RCC与癌旁组织中miR-660的表达,qPCR检测40例RCC与癌旁组织中miR-660的表达。结果:MPSS结果显示,RCC组织中283个miRNAs表达下调,187个表达上调,其中miR-660在RCC组织中表达量是癌旁组织的17.5%。RT-PCR初步验证了此结果;qPCR验证结果显示,40例RCC组织中33例miR-660表达显著低于癌旁组织,RCC组织中miR-660平均表达量是癌旁的19.5%。结论:人RCC组织中283个miRNAs表达下调、187个上调。miR-660在RCC组织中低表达,有可能成为RCC诊断及治疗的新靶点。
OBJECTIVE: To screen differentially expressed microRNAs (miRNAs) in human renal cell carcinoma (RCC) and adjacent tissues by massively parallel signature sequencing (MPSS) and to validate the expression of miRs, one of the differentially expressed miRNAs -660 in human RCCs. Methods: MPSS was used to detect the expression of miRNAs in 10 cases of RCC tissues and corresponding paracancerous tissues, and to screen differentially expressed miRNAs in RCC tissues. The expression of miR-660 in 5 cases of RCC and adjacent non-cancerous tissues was detected by RT-PCR. The expression of miR-660 in 40 cases of RCC and paracancerous tissues was detected by qPCR. Results: The results of MPSS showed that 283 miRNAs were down-regulated and 187 were up-regulated in RCC tissues. The expression of miR-660 in RCC tissues was 17.5% of that in paracancerous tissues. The results of qPCR showed that the expression of miR-660 in 33 RCC tissues was significantly lower than that in paracancer tissues, and the average expression of miR-660 in RCC tissues was 19.5% of that in adjacent tissues. Conclusion: 283 miRNAs were down-regulated and 187 up-regulated in human RCC tissues. Low expression of miR-660 in RCC tissues may become a new target for the diagnosis and treatment of RCC.