MiR-134通过靶向调节KRAS抑制786-0肾透明细胞癌细胞上皮间质转化过程  

MiR-134 inhibits epithelial-to-mesenchymal transition by targeting KRAS in 786-0 renal cell carcinoma cell

作　　者：鲍美玲[1] 钱健[1] 张少波[1] 李双[1] 张磊[1] 赵睿哲[1] 秦超[1] 邵鹏飞[1] 殷长军[1] 

机构地区：[1]南京医科大学第一附属医院泌尿外科,江苏南京210029

出　　处：《现代泌尿外科杂志》2014年第6期393-398,共6页Journal of Modern Urology

基　　金：国家自然科学基金(No.81171963;8201571);江苏省临床医学科技专项(No.BL2012027)

摘　　要：目的探索miR-134在肾癌组织中的表达并研究其对肾透明细胞癌细胞786-0上皮间质转化的影响。方法采用逆转录-聚合酶链反应(RT-PCR)检测miR-134在20对肾癌组织和786-0细胞中的表达情况;miR-134模拟物转染786-0后,划痕实验和Transwell实验分别检测肾癌细胞迁移和侵袭能力的改变情况,Western blot检测细胞中靶蛋白KRAS及其相关信号通路蛋白表达情况;双荧光素酶报告基因检测miR-134与KRAS 3′-UTR结合情况。结果 MiR-134在20对肾癌组织和786-0细胞中明显低表达(P<0.01);miR-134模拟物转染后能够抑制肿瘤细胞侵袭(P<0.01)和迁移的能力(P<0.05),显著降低肿瘤细胞中KRAS蛋白的水平(P<0.05)并抑制RAS/MAPK/ERK通路中关键蛋白p-ERK的表达(P<0.05);双荧光素酶报告基因显示miR-134能与KRAS 3′-UTR结合,显著降低荧光值(P<0.05)。结论 MiR-134在肾透明细胞癌中显著低表达,能通过靶向抑制KRAS表达调控下游RAS/MAPK/ERK通路活性,阻滞786-0细胞上皮间质转化过程,抑制肿瘤细胞的侵袭和转移。Objective To investigate the expression of miR-134 in human renal cell carcinoma （RCC） and to explore its function in 786-0 RCC cells.Methods Real-time PCR was used to analyze the expression of miR-134 in human RCC samples and 786-0 cells.Wound healing assay was carried out to detect the ability of migration in transfected 786-0 cells; the ability of invasion was detected by transwell invasion assay.Western blot was adopted to identify the potential signaling pathways and the alterations of the markers in epithelial-to-mesenchymal transition （EMT） process which affected RCC metastasis by miR-134.A luciferase reporter assay was performed to confirm whether KRAS was a direct target of miR-134.Results MiR-134 was found downregulated in RCC samples and 786-0 RCC cells （P〈0.01）.Overexpression of miR-134 suppressed migration （P〈 0.05） and invasion （P〈0.01） by blocking EMT in 786-0 cells.KRAS was identified as a novel target of miR-134 and miR-134 might act as a tumor suppressor via KRAS-related MAPK/ERK pathway signaling.Conclusions MiR-134 was downregulated in RCC and could inhibit EMT by targeting KRAS in 786-0 cells.

关 键 词：miR-134 微小RNA 上皮间质转化 KRAS 肾癌 

分 类 号：R733[医药卫生—肿瘤]