大规模平行测序技术筛选人肾细胞癌组织中差异表达microRNAs及其验证

Massively parallel signature sequencing in screening differentially expressed microRNAs in human renal cell carcinoma tissues and their identification

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 tumor and the corresponding paracancerous tissues of renal cell carcinoma（RCC） by massively parallel signature sequencing（MPSS）,and to identify the expression of miR-660 in RCC.Methods： The expression of miRNAs in tumor and the corresponding paracancerous tissues of 10 RCC patients was examined by MPSS,and the differentially expressed miRNAs were screened.Expression of miR-660 in tumor tissues of 5 patients and the paracancerous tissues of 40 RCC patients were further examined by RT-PCR and qPCR,respectively.Results： The MPSS results showed that 283 miRNAs were up-regulated and 187 were down-regulated in RCC tissues,and the expression level of miR-660 in RCC tissues was only 19.5% of that in the paracancerous tissues.The MPSS results were primarily confirmed by RT-PCR,and qPCR results further showed that miR-660 expression in 34 of 40 RCC tissues was significantly lower than that in the paracancerous tissues,with the average level being only 11.7% of that in the paracancerous tissues.Conclusion： A total of 283 miRNAs are up-regulated and 187 down-regulated in RCC tissues;and miR-660 is lowly expressed in RCC,which may be a new target for the diagnosis and treatment of RCC.