miR-34a通过调控YY1抑制肾癌细胞的增殖及侵袭

目的探讨miR-34a在肾癌组织中表达、miR-34a对人肾癌ACHN细胞增殖和侵袭的影响及调控机制。方法实时聚合酶链反应(PCR)检测miR-34a在20例肾癌及癌旁组织中的表达;采用脂质体转染法将miR-34a mimics转染入肾癌ACHN细胞中,试验分空白对照组、阴性对照组、miR-34a mimics转染组。实时PCR检测转染24 h后miR-34a表达量;噻唑蓝(MTT)试验检测细胞增殖;流式细胞术检测细胞周期;Transwell及Matrigel检测细胞侵袭能力;实时PCR和蛋白质印迹(Western blot)技术检测转染后转录因子YY1表达水平。结果与癌旁组织相比,20例癌组织中miR-34a平均表达量为1.06±0.67,显著低于癌旁组织(1.62±0.83,P<0.01);转染后24 h,miR-34a mimics转染组的miR-34a表达量为157.04±13.01,较阴性对照组显著上调(P<0.01);miR-34a mimics转染组细胞增殖能力显著降低(P<0.01),细胞生长被阻滞在G0/G1期;细胞侵袭能力明显减弱(P<0.01);转染miR-34a mimics后YY1基因在mRNA表达无明显变化(P>0.05),蛋白水平下调。结论 miR-34a在肾癌中低表达,可能与肾癌的发生有关;miR-34a调控YY1的表达可能是抑制肾癌细胞生物活性的重要机制之一。

The inhibition of cell proliferation and invasion through miR-34a regulating YY1 in human renal carcinoma cell

Objective To investigate miR-34a expression in renal carcinoma cell and the inhibitory effect and regulating mechanism of miR-34a on the proliferation and invasion in human renal carcinoma ACHN cells.Methods The expressions of miR-34a in cancer and pericancerous tissues were detected by real-time polymerase chain reaction（PCR）.ACHN cells were transfected with miR-34a mimics and there set blank control group,negative control group and miR-34a mimics group.The expression of miR-34a was measured by real-time PCR 24 h after transfection.The cell proliferation and cell cycle were determined by 5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide（MTT） and flow cytometry.The invasive ability was examined by Transwell and Matrigel invasive assays.The mRNA and protein levels of YY1 were detected by real-time PCR and Western blot.Results The relative expression of miR-34a in cancer tissues（1.06±0.67） was significantly lower than that in pericancerous tissues（1.62±0.83,P〈0.01）.After transfection 24 h,an increase expression of miR-34a was noted in miR-34a mimics group（157.04±13.01） comparing with negative control group（P〈0.01）.Overexpression of miR-34a significantly inhibited the cell proliferation（P〈0.01）.The cell cycle was arrested at G0/G1 phases.Transwell and Matrigel invasive assays showed that the invasive ability of cells was significantly suppressed after transfection（P〈0.01）.YY1 gene had no mRNA expressions after transfection（P〉0.05）.Conclusions The expression of miR-34a is low in renal carcinoma,and it may be correlated with tumorigenesis,partially through regulating YY1.