miR-646高表达抑制EGFR/Akt通路及肺癌细胞增殖扩散的研究

目的 探讨微小RNA(microRNA,miR)-646对EGFR/Akt通路的作用及对肺癌A549细胞增殖扩散的影响及相关的作用机制.方法 利用Lipo2000将miR-646转染到肺癌A549细胞中,采用实时荧光定量PCR(real time PCR,RT-PCR)法检测转染后各组细胞中miR-646的表达;CCK-8法检测细胞增殖;划痕和Transwell小室分别检测细胞的迁移和侵袭能力;Western blotting法检测EGFR/Akt通路中各蛋白的变化水平.结果 RT-PCR结果显示,转染miR-646组其miR-646表达水平显著高于阴性对照组和空白对照组(P＜0.05),A549细胞转染miR-646后,细胞增殖和扩散能力均显著降低(P＜0.05);Western blotting结果显示,转染miR-646组中p-EGFR和p-Akt的蛋白表达水平均明显降低(P＜0.05).结论 miR-646可显著降低肺癌A549细胞中p-EGFR、p-Akt的蛋白水平,进而抑制细胞的增殖和扩散能力.

Inhibition of EGFR/Akt Pathway Through miR-646 and the Proliferation and Spread of Lung Cancer Cell

Objective To investigate the effects of microRNA (miRNA)-646 on EGFR/Akt signal pathway and the proliferation and spread of human lung cancer cell lines.Methods The A549 cells were transfected with miR-646 by Lipo2000.The expression of miR-646 was detected by real-time quantitative PCR(RT-PCR).The cell proliferation was detected by CCK-8 assay.The cell migration and invasion abilities wereexamined by wound healing and transwell methods respectively.And the protein levels of p-EGFR,EGFR,p-Akt,Akt were determined by Western blotting technology.Results The expression level of miR-646 was significantly increased in the A549 cells Compared with negative control (NC) group and control group (P ＜ 0.05).The proliferation of A549 cells was decreased significantly after transfected with miR-646 (P ＜ 0.05).The migration and invasion abilities of the lung cancer cells were also significantly inhibited.The protein levels of p-EGFR,p-Akt were significantly down-regulated compared with NC group and control group (P ＜ 0.05).Conclusion Over-expression of miR-646 significantly inhibits the proliferation and spreadability of human lung cancer A549 cells by down-regulation of EGFR/Akt pathway.