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作 者:陈鹏[1] 刘博雅[1] 汪健[1] 赵亚超[1] 卢强[1] 邓迎春[1] 张志培[1] 李小飞[1]
出 处:《现代生物医学进展》2010年第22期4201-4203,4241,共4页Progress in Modern Biomedicine
基 金:国家自然科学基金(NO.30973379);陕西省科学技术研究发展计划基金(NO.2008K09-01)项目资助
摘 要: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.
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