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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

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【作者】 翟庆娜周亮余振东唐爱发王勇桂耀庭

【Author】 ZHAI Qing-na1,2,ZHOU Liang3,YU Zhen-dong1,TANG Ai-fa3,WANG Yong3,GUI Yao-ting3△(1.Department of Clinical Laboratory,Shenzhen Hospital of Peking University,Shenzhen 518036,Guangdong,China;2.Department of Clinical Laboratory Diagnostics,Shantou University Medical College,Shantou 515031,Guangdong,China;3.Key Laboratory of Male Reproductive Medicine and Genetics in Guangdong,Shenzhen Hospital of Peking University,Shenzhen 518036,Guangdong,China)

【机构】 北京大学深圳医院检验科汕头大学医学院临床检验诊断学教研室北京大学深圳医院男性生殖与遗传学省重点实验室

【摘要】 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。

【Abstract】 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.

【基金】 国家自然科学基金资助项目(No.30900817);深港创新圈项目资助(No.ZYB200907080110A)~~
  • 【文献出处】 中国肿瘤生物治疗杂志 ,Chinese Journal of Cancer Biotherapy , 编辑部邮箱 ,2011年03期
  • 【分类号】R737.11
  • 【被引频次】3
  • 【下载频次】204
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