TRPC1对人肾癌细胞A498增殖和迁移的影响  

Effect of TRPC1 on proliferation and migration of human renal carcinoma cell A498

作　　者：阮健[1] 蔡红兵[1] 罗兰[1] 宋姗姗[1] 肖雅娟[1] 张军一[1] 

机构地区：[1]南方医科大学中西医结合医院,广东广州510315

出　　处：《热带医学杂志》2016年第7期825-828,F0004,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(81401874);南方医科大学中西医结合医院院长基金(1201302009);广东省自然科学基金(2015A030313250)

摘　　要：目的探讨瞬时受体电位C1(TRPC1)对人肾癌细胞A498细胞增殖和迁移的影响。方法利用荧光定量PCR检测人肾癌细胞A498、786-O和GRC-1中TRPC1的表达水平。设计并合成靶向TRPC1的特异性sh RNA,通过脂质体转染法转染TRPC1表达最高的人肾癌细胞A498以构建稳定低表达TRPC1细胞株,通过免疫印迹法和荧光定量PCR来验证sh RNA的干扰效率;通过MTT比色法检测干扰后细胞的增殖能力;通过Transwell小室测细胞迁移情况;免疫印迹法检测PI3K和AKT的表达水平。结果在3株肾癌细胞中A498的TRPC1表达量最高;特异性靶向TRPC1的sh RNA能够有效下调A498细胞中TRPC1的表达;特异性sh RNA干扰组细胞较其他两组细胞增殖显著减慢(F=42.31,P<0.01);A498/sh RNA组的迁移细胞数显著低于A498组和A498/Con组(F=24.52,P<0.01);A498/sh RNA组细胞的PI3K和AKT蛋白的表达较A498组和A498/Con组显著下调。结论采用RNA干扰技术能够有效沉默A498细胞的TRPC1基因,并致使其细胞增殖能力下降以及细胞迁移能力减弱,其中可能的机制为通过调节细胞因子PI3K来调控人肾癌细胞的增殖和迁移。提示TRPC1在肾癌的发生发展中起着较为重要作用,TRPC1可能成为治疗肾癌的新靶点。Objective To investigate the effect of down-regulation of TRPC1 on the proliferation and migration of human renal carcinoma cell A498. Methods shRNA targeting TRPC1 was designed and synthesized, and then transfected into the A498 cells via Lipofectamine 2000 mediation. The proliferation and migration ability of A498 were detected by using MTT assay and transwell assay. Expression of PI3K and AKT were detected by using western blot. Results TRPC1-targeted sh RNA could down-regulate the TRPC1 expression of A498. MTT assay showed a significant decrease of the proliferation of the sh RNA interference group as compared with blank group and control group（P〈0.01）. Compared with control group, migrated cell were significantly lower in sh RNA interference group（P〈0.01）. In addition, the expression of PI3K and AKT were decreased in sh RNA interference group. Conclusion Down-regulation of TRPC1 can induce inhibition of proliferation and migration in human renal carcinoma cells A498 via regulating PI3K. It could be regarded as a novel target for clinical diagnosis and gene therapy for renal cancer.

关 键 词：瞬时受体电位C1 细胞增殖 细胞迁移 肾癌 

分 类 号：R737.11[医药卫生—肿瘤]