miR-34a通过调控YY1抑制肾癌细胞的增殖及侵袭

检验医学 ›› 2012, Vol. 27 ›› Issue (8): 635-640.

miR-34a通过调控YY1抑制肾癌细胞的增殖及侵袭

王明丽¹,李智²,徐晋豫¹,许闪闪¹,翁文浩¹

同济大学附属上海市第十人民医院检验科，上海 200072

收稿日期:2012-03-09 出版日期:2012-08-30 发布日期:2012-08-08
通讯作者: 李　智，联系电话：021-66306831。
作者简介:王明丽，女，1984年生，技师，主要从事分子生物学检验工作。

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

Department of Clinical Laboratory，the Tenth People′s Hospital of Tongji University，Shanghai 200072，China

Received:2012-03-09 Online:2012-08-30 Published:2012-08-08
Contact: zhi LI

摘要/Abstract

摘要： 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。

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关键词: miR-34a, YY1, 细胞增殖, 侵袭, 肾癌

Abstract: 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.

王明丽;李　智;徐晋豫;翁文浩;许闪闪. miR-34a通过调控YY1抑制肾癌细胞的增殖及侵袭[J]. 检验医学, 2012, 27(8): 635-640.

WANG Mingli;LI Zhi;XU Jinyu;WENG Wenhao;XU Shanshan. The inhibition of cell proliferation and invasion through miR-34a regulating YY1 in human renal carcinoma cell[J]. , 2012, 27(8): 635-640.

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