α-硫辛酸下调Grb2抑制A549细胞增殖的研究  

Alpha Lipoic Acid Inhibits A549 Cell Proliferation through the Repression of Grb2

作　　者：文娅 李贝 邓雪松[4] 林云涛 陆菁潇 吕国庆[3] WEN Ya;LI Bei;DENG Xue-song;LIN Yun-tao;LU Jing-xiao;LV Guo-qing(Institute of Translational Medicine, Shenzhen Second People's Hospital, Shenzhen, Guangdong,518035, China;Department of Gastrointestinal Surgery, Peking University Shenzhen Hospital, Shenzhen, Guangdong,518036, China;Department of Hepatobiliary Surgery, Shenzhen Second People's Hospital, Shenzhen, Guangdong,518035, China;Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital, Shenzhen, Guangdong,518036, China;Dongguan HEC Technology Development and Research Co., Ltd, Dongguan, Guangdong,523843, China)

机构地区：[1]深圳市第二人民医院转化医学研究院,广东深圳518035 [2]东莞东阳光科研发有限公司,广东东莞523843 [3]北京大学深圳医院胃肠外科,广东深圳518036 [4]深圳市第二人民医院肝胆外科,广东深圳518035 [5]北京大学深圳医院口腔科,广东深圳518036

出　　处：《现代生物医学进展》2018年第21期4012-4016,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81672813);人事部中国博士后科学基金项目(2016M602594);深圳市科技计划2016年基础研究项目(JCYJ20160425105945739);深圳市"三名工程"(SZSM201612051)

摘　　要：目的:探讨α-LA对A549细胞增殖的机制以及Grb2在其中发挥的作用。方法:利用CCK-8和流式细胞术检测α-LA对细胞增殖的影响;实时定量PCR和Western Blot检测细胞中Grb2表达的变化;利用向细胞中转染siGrb2及过表达Grb2载体检测细胞中Grb2表达水平差异对细胞增殖的影响。结果:α-LA处理A549 24 h后,对照组和α-LA处理组中位于G_0/G_1期细胞的比率由40.60%上升至57.80%;而位于S期细胞的比率由45.96%下降至39.01%;与细胞G1/S期的转换相关的调节蛋白CDK2/4/6,cyclin D3和E1的表达也随之发生了相应的改变,α-LA通过抑制细胞G_1/S期的转换而抑制细胞增殖。与亲本细胞相比,Grb2在α-LA处理A549细胞中的转录和蛋白表达水平均显著降低,干扰细胞中Grb2表达能显著抑制A549增殖;而过表达Grb2可阻断α-LA诱导的细胞增殖抑制。结论:α-LA具有抑制A549细胞增殖的作用,Grb2是α-LA发挥抑细胞增殖效应的重要介导因子。Objective: To explore the roles of growth factor receptor-bound protein 2 (Grb2) in Alpha lipoic acid inhibits cell proliferation process. Methods: The CCK-8 assay and flow cytometry were used to assess cell proliferation in A549 cell lines after α-LA treatment. The expression of Grb2, cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, Cyclin D3, Cyclin E1 was measured by western blotting. Grb2 levels were restored in α-LA-treated cells by transfection of a plasmid carrying Grb2 and were reduced in A549 cells via specific siRNA knockdown. Results: CCK8 assay and flow cytometry were shown that α-LA dramatically decreased A549 cell proliferation. In control cells cultured without α-LA, the cells in the G 1 and S phases were 40.60% and 45.96%, respectively. In α-LA treated group, the cells in the G 1 and S phases were 57.80% and 39.01%, respectively, at 24 h Western Blot analysis indicated that α-LA decreased the levels of CDK2/4/6, cyclin D3 and E1, events that were associated with the inhibition of the G 1 /S-phase transition. The level of Grb2 was downregulating in α-LA treated cells; in contrast, Grb2 overexpression significantly prevented α-LA-induced decreases in cell growth in vitro. Conclusion: These findings provide the first evidence that α-LA could inhibit cell proliferation and Grb2 is involved in this process.

关 键 词：Α-硫辛酸 非小细胞肺癌 细胞增殖 GRB2 

分 类 号：R-33[医药卫生] R734.2