NOK过表达对肺腺癌细胞A549增殖的影响

Effect of NOK Over-expression on Proliferation of Lung Adenocarcinoma Cell A549

目的:本研究通过建立稳定过表达NOK基因的A549-NOK细胞系观察其增殖变化,探讨NOK基因对A549增殖的影响。方法:运用电穿孔仪将pcDNA3.1-NOK质粒转染人肺癌A549细胞,筛选出稳定过表达NOK基因的单克隆株A549-NOK。RT-PCR检测转染前后细胞内NOK基因的表达效果。通过流式细胞术检测细胞周期,MTT法绘制细胞生长曲线。结果:RT-PCR显示A549-NOK细胞中NOK基因表达量明显增高。流式细胞术显示A549-NOK与A549、空质粒对照组细胞相比,S期细胞增多(51.7±2.2 vs 43.4±1.5,45.7±1.4,均P<0.05);细胞增殖指数升高(69.4±1.1vs 58.4±1.3,57.9±1.4,均P<0.05);MTT结果显示A549-NOK细胞较A549和空质粒对照组细胞生长曲线上移。结论:NOK可促进A549细胞的增殖,可能在肺癌的发生、发展中发挥重要作用。

Objective：To establish the stable expression of NOK in lung adenocarcinoma cell line A549 and to investigate the infection of NOK high expression to the proliferation of A549.Methods：pcDNA3.1-NOK Plasmid was transferred into A549 cells with electroporation apparatus.The cloned strain A549-NOK with stable expression of NOK was screened and tested by RT-PCR.The cell cycle was detected by flow cytometry and the cell growth curve was determined by MTT.Results：There was the stable high expression level of NOK in A549-NOK cells.The number of A549-NOK cells in S stage was larger than that of A549 cells and blank contriol（51.7±2.2 vs 43.4±1.5,45.7±1.4,both P〈0.05）;The proliferation index（PI） in A549-NOK was higher than that in A549 and blank contriol（69.4±1.1 vs 58.4±1.3,57.9±1.4,both P〈0.05）.The growth curve of A549-NOK was steeper than that of A549 and blank contriol.Conclusions：NOK could improve the proliferation of the A549 cell and might play an important role in the development of human lung cancer.