防己诺林碱对肺癌H1299和A549细胞的作用及其分子机制研究  

Effect of tetrandrine on lung cancer H1299 cells and A549 cells and its molecular mechanism

作　　者：陈澜涛[1] 颜巍[1] 刘静蕾[1] 姚磊[1] CHEN Lantao;YAN We;LIU Jinglei;YAO Lei(Harbin Medical University Cancer Hospital,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学附属肿瘤医院,哈尔滨150081

出　　处：《实用肿瘤学杂志》2020年第2期126-132,共7页Practical Oncology Journal

基　　金：2018年黑龙江省中医药科研项目(编号:ZHY18-159);黑龙江省卫生计生委科研课题(编号:2018240)。

摘　　要：目的探究防己诺林碱在肺癌中的作用及其分子机制。方法MTS法检测经10、15、20、30、40和60μM/L防己诺林碱处理的肺癌H1299和A549细胞的增殖情况,通过Caspase3、Caspase8和Caspase9活性检测试剂盒和TUNEL试剂盒检测细胞凋亡情况。Western blot检测MAPK信号通路(p-Akt、PI3K、mTOR和Akt蛋白)、增殖和转移(MMP-2、MMP-9、PCNA、Cyclin D1和P21蛋白)、周期和凋亡(Cyclin D2、Bcl2、MCL-1、Bax和p53蛋白)相关蛋白的表达情况,通过Real-time PCR检测PI3k、mTOR、MMP-2、MMP-9、PCNA、Cyclin D1、Cyclin D2、MCL-1、Bax、Bcl2和TP53基因表达情况。结果防己诺林碱会抑制肺癌H1299和A549细胞的增殖,诱导细胞凋亡,增加Caspase3、Caspase8和Caspase9酶活性。经防己诺林碱处理后,PI3K、p-Akt、mTOR、MMP-2、MMP-9、PCNA、Cyclin D1、Cyclin D2和Bcl2蛋白表达降低,P21、MCL-1、Bax和p53蛋白表达升高,并且表现为剂量依赖性,但对Akt蛋白表达无影响;PI3K、mTOR、MMP-2、MMP-9、PCNA、Cyclin D1、Cyclin D2和Bcl2基因表达降低,TP53、MCL-1和Bax基因表达增加。结论防己诺林碱通过调控MAPK信号通路、增殖和转移、周期和凋亡相关蛋白和基因的表达,从而抑制细胞增殖,诱导细胞凋亡。Objective The objective of this study was to explore the role of tetrandrine in lung cancer and its molecular mechanism.Methods The MTS method was used to detect the proliferation of lung cancer H1299 cells and A549 cells treated with tetrandrine at 10,15,20,30,40 and 60μM/L,and then the activities of Caspase3,Caspase8 and Caspase9 were detected by kits,and apoptosis was examined by TUNEL kit.In order to study the mechanism of tetrandrine,Western blot was used to detect the expression of MAPK signaling pathways(p-Akt,PI3K,mTOR and Akt proteins),proliferation and metastasis(MMP-2,MMP-9,PCNA,Cyclin D1 and P21 proteins),cell cycle and apoptosis related proteins(Cyclin D2,Bcl-2,MCL-1,Bax and p53 proteins).Finally,Real-time PCR was used to detect the mRNA expression of PI3k,mTOR,MMP-2,MMP-9,PCNA,Cyclin D1,Cyclin D2,MCL-1,Bax,Bcl-2 and TP53.Results Tetrandrine could inhibit the proliferation of lung cancer H1299 and A549 cells,induce apoptosis,and increase the activities of Caspase3,Caspase8 and Caspase9 in H1299 cells and A549 cells.After treatment with tetrandrine,the expression of PI3K,p-Akt,mTOR,MMP-2,MMP-9,PCNA,Cyclin D1,Cyclin D2and Bcl-2 proteins were reduced,and the expression of P21,MCL-1,Bax and p53 proteins were increased,showing a dose-dependent manner,but no effect on the expression of Akt protein.The mRNA expression of PI3K,mTOR,MMP-2,MMP-9,PCNA,Cyclin D1,Cyclin D2 and Bcl-2 was decreased,and the mRNA expression of TP53,MCL-1 and Bax was increased in H1299 cells and A549 cells.Conclusion Tetrandrine inhibits cell proliferation and induces apoptosis by regulating the MAPK signaling pathway,proliferation and metastasis,cell cycle and apoptosis related proteins and genes expression.

关 键 词：防己诺林碱 肺癌 分子机制 

分 类 号：R734.2[医药卫生—肿瘤]