GPRC5A抑制肺癌细胞A549增殖和迁移及与EGFR的相关性研究

GPRC5A inhibits proliferation and migtation of lung adenocarcinoma A549 cells and its relation to EGFR

目的探讨G蛋白偶联受体C家族5A(GPRC5A)基因对肺癌细胞A549增殖和迁移能力的影响及其与表皮生长因子受体(EGFR)的相关性。方法构建GPRC5A基因真核表达质粒pLenti6.3-MCS-GPRC5A,通过脂质体法转染A549细胞,经过新霉素筛选获得阳性单克隆细胞。用qRT-PCR和Western blot检测转染前后GPRC5A、EGFR基因的m RNA和蛋白表达,CCK-8法检测细胞增殖能力的变化,Transwell法检测细胞迁移能力的变化。结果真核表达质粒转染A549细胞后,GPRC5A m RNA表达上调,蛋白表达水平升高,EGFR m RNA表达下降,蛋白表达水平降低。高表达GPRC5A后,细胞增殖和迁移能力显著减弱。结论GPRC5A可能与EGFR结合,抑制EGFR通路的过度激活,从而影响了A549细胞的增殖和迁移能力。

Objective To investigate the effect of GPRC5A gene on the proliferation, migration of lung adenocarcinoma A549 cells and its relation to EGFR. Methods A recombinant eukaryotic expression plasmid pLenti6.3-MCS-GPRC5A was constructed and transfected into A549 cells by lipofectamine 2000. Positive monoclone was screened by G418. Quantitative RT-PCR and Western blot assays were used to detect the mRNA and protein expression of GPRC5A and EGFR, respectively. The effects of GPRC5A on A549 cell proliferation and migration were determined with CCK8 and Transwell assays, respectively. Results The GPRC5A mRNA and protein were overexpression in transfected A549 cells, but the expression of EGFR mRNA and protein was down-regulated. Up-regulation of GPRC5A led to inhibition of the proliferation and migration of A549 cells. Conclusion GPRC5A may bind to EGFR to inhibit the excessive activation of EGFR pathway, which affect the proliferation and migration of A549 cells.