黄芪甲苷通过调节细胞自噬增强贝伐单抗在肺癌细胞系A549中抗肿瘤作用机制的研究  

Astragaloside enhanced the anti-tumor effect of bevacizumab by regulating cell autophagy in lung cancer cell line A549

作　　者：庞歆桥 王婧[1] 姚婷[2] 龚明涛[2] 马妮娜[1] 

机构地区：[1]首都医科大学附属北京友谊医院肿瘤科,北京100050 [2]首都医科大学基础医学院医学遗传学系,北京100069

出　　处：《临床与病理杂志》2016年第3期220-225,共6页Journal of Clinical and Pathological Research

基　　金：首都医科大学基础-临床合作课题资助项目(2014-2015);北京市医院管理局"青苗"计划专项经费资助(QML20150107);北京市中医药科技发展资金项目资助(QN2015-08);首都医科大学附属北京友谊医院院启动基金资助(yyqdkt2014-10);首都卫生发展科研专项基金资助(2016)~~

摘　　要：目的:验证黄芪甲苷可以通过调节细胞自噬增强贝伐单抗对肺癌细胞系A549细胞增殖能力的影响,讨论黄芪甲苷在A549中的抗肿瘤作用机制。方法:MTT方法检测黄芪甲苷单药以及与贝伐单抗联合应用对A549细胞增殖能力的影响。Western blot方法分别检测黄芪甲苷以及贝伐单抗处理A549后自噬相关蛋白P62和LC3的表达水平。RT-PCR方法检测黄芪甲苷处理后A549细胞内侵袭相关基因MMP-2和MMP-9的表达情况。ECIS方法检测黄芪甲苷处理后A549细胞黏附与迁移能力的变化。结果:MTT检测结果显示,100 mg/L的黄芪甲苷与750 mg/L的贝伐单抗联合应用对A549细胞的活力有显著的抑制作用,两药联合应用组与对照组相比差异有统计学意义,P值为0.0009。Wester n blot检测结果显示,黄芪甲苷能够通过影响自噬通路P62和LC3的表达从而抑制自噬的发生。RT-PCR方法显示黄芪甲苷能显著抑制侵袭相关基因MMP-2和MMP-9的表达,黄芪甲苷组与对照组相比差异有统计学意义,P值分别为0.0001和0.001。ECIS结果显示黄芪甲苷能够增强A549细胞的黏附能力并抑制其转移。结论:体外研究结果显示黄芪甲苷能够增强A549细胞的黏附能力并抑制其转移和侵袭,这一过程是通过抑制细胞自噬的发生,而贝伐单抗有较弱的促进自噬发生的作用,当两者联合时较单用贝伐单抗更强的抗肿瘤细胞增殖的作用。Objective：To investigate the antitumor mechanism of astragaloside（AST） with bevacizumab in the lung cancer cell line A549 by the cell proliferation,adhesion,migration and invasion.Methods：The cell proliferation of lung cancer cell line A549 intervened by astragaloside with or without bevacizumab was detected by 4-methylteerazolium（MT）.the expressive change for protein of P62 and LC3 in A549 cells was detected by Western blot.the cell invasion of lung cancer cell line A549 was detected by RT-PCR through the expression of the gene MMP-2 and MMP-9.Results：MT showed that the best concentration in A549 was 100 mg/L astragaloside combined with 750 mg/L bevacizumab which had signii cantly inhibition of cell proliferation in A549 cells.Western blot showed that astragaloside can affect the expression of P62 and LC3 to suppress the occurrence of autophagy（P=0.0009）.RT-PCR showed that astragaloside can inhibit the expression of invasion related gene MMP-2 and MMP-9（P=0.0001,0.001）.ECIS results showed that astragaloside can enhance the adhesion and suppress the migration of A549.Conclusion：In vitro studies showed that astragaloside can inhibit autophagy and astragaloside with bevacizumab has stronger antitumor effect than bevacizumab without astragaloside.

关 键 词：黄芪甲苷 肺癌 自噬 贝伐单抗 

分 类 号：R734.2[医药卫生—肿瘤]