黄连素联合顺铂诱导人肺癌耐药A549/DDP细胞凋亡的实验研究  

An Experimental Study on Apoptosis of A549/DDP Cells Induced by Berberine Combined with Cisplatin

作　　者：曾玲芳 王亿君 吴依辰 章慧[2] ZENG Lingfang;WANG Yijun;WU Yichen;ZHANG Hui(Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Cancer Hospital, Changsha, Hunan 410006, China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省肿瘤医院,湖南长沙410006

出　　处：《湖南中医药大学学报》2018年第5期493-498,共6页Journal of Hunan University of Chinese Medicine

基　　金：湖南省科学技术厅资助项目(2014FJ3133)

摘　　要：目的观察黄连素联合顺铂对A549/DDP细胞凋亡的影响并探讨其相关机制。方法以A549/DDP细胞为研究对象,分别加入12μg/m L的顺铂、浓度为20μmol/L、40μmol/L、80μmol/L的黄连素+12μg/m L的顺铂,设置空白对照孔(PBS孔),分别用药物干预24 h、48 h、72 h,采用MTT法检测细胞增殖活性;各组用药干预48 h后,流式细胞仪检测细胞凋亡,Western blot法检测Caspase-3、Caspase-9、Bax、Bcl-2的蛋白表达情况,Real-time PCR法检测Caspase-3、Caspase-9、Bax、Bcl-2的m RNA表达水平。结果黄连素联合顺铂能够抑制A549/DDP细胞增殖(P<0.01),并具有一定的时间-浓度依赖性。黄连素联合顺铂能够诱导耐药细胞A549/DDP凋亡(P<0.01),且呈一定的浓度依赖性。不同浓度的黄连素联合顺铂使促凋亡相关基因Caspase-3、Caspase-9、Bax在蛋白和m RNA水平表达上调(P<0.05),而抗凋亡基因Bcl-2在蛋白和m RNA水平表达下调(P<0.05)。结论黄连素联合顺铂抑制A549/DDP细胞增殖和促凋亡的分子机制可能是上调促凋亡相关基因Caspase-3、Caspase-9、Bax的表达,下调抗凋亡基因Bcl-2的表达。Objective To observe the effect of berberine combined with cisplatin on cell apoptosis of A549/DDP and investgate its related mechanism. Methods Human lung cancer resistance A549/DDP cells, as the object of study, were added with 12 μg/m L cisplatin, and 20 μmol/L, 40 μmol/L, 80 μmol/L berberine combined with 12 μg/m L cisplatin,respectively, blank control wells（PBS wells） were set up, they were all treated with drugs for 24 h, 48 h and 72 h. The cell proliferation was evaluated by MTT assay. After intervention with drugs for 48 h, cell apoptosis was determined by using the flow cytometry instrument. The protein and m RNA levels of Caspase-3, Caspase-9, Bax, Bcl-2 were measured by Real-time PCR and western blotting analysis, respectively. Results Berberine combined with cisplatin could inhibit the growth of A549/DDP cells（P〈0.01） and promote apoptosis of A549/DDP cells（P〈0.01）, with a certain time-concentration dependence.Different concentrations of berberine combined with cisplatin increased the levels of protein and m RNA levels of apoptosis genes Caspase-3, Caspase-9, Bax（P〈0.05）, and down-regulated the protein and m RNA levels of anti-apoptosis gene Bcl-2（P〈0.05）.Conclusion The molecular mechanism of berberine combined with cisplatin in inhibiting proliferation and promoting apoptosis of A549/DDP cells may be related to up-regulating the expression of apoptosis related proteins Caspase-3, Caspase-9 and Bax, downregulating the expression of anti-apoptosis protein Bcl-2.

关 键 词：黄连素 顺铂 肺癌耐药 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学] R734.2[医药卫生—中医学]