双硫仑联合铜对人肾上腺皮质癌H295R细胞增殖和凋亡的影响及机制研究  

Effects and mechanisms of disulfiram combined with copper on the proliferation and apoptosis of human adrenocortical carcinoma H295R cells

作　　者：邢增苗 苏婷婷 匡雅琪 黄振兴[1] 杨海燕[1] 巫丽丽[1] 罗佐杰[1] Xing Zengmiao;Su Tingting;Kuang Yaqi;Huang Zhenxing;Yang Haiyan;Wu Lili;Luo Zuojie(Department of Endocrinology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院内分泌科

出　　处：《广西医科大学学报》2019年第5期680-685,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81060220)

摘　　要：目的:观察双硫仑(DSF)联合铜(Cu)对人肾上腺皮质癌细胞H295R的抑制增殖作用与诱导细胞凋亡的影响,并探讨其相关分子机制。方法:体外培养H295R细胞,设立空白对照组(Con组)、Cu组(加入0.5μmol/LCu^2+)及不同浓度的DSF/Cu组(分别加入0.1μmol/L、0.2μmol/L、0.4μmol/L、0.8μmol/L、1.6μmol/LDSF及0.5μmol/LCu^2+),采用噻唑蓝(MTT)法检测药物干预12h、24h、48h后对细胞的增殖抑制作用;流式细胞术检测细胞凋亡率;Westernblotting法检测细胞Bcl-2和Bax蛋白的表达。结果:DSF/Cu对H295R细胞有显著抑制增殖作用(P<0.05),且呈时间及浓度依赖性;而Cu组与Con组细胞增殖抑制率比较,差异无统计学意义(P>0.05)。DSF/Cu干预24h后对细胞的半数抑制浓度(IC50)为0.27μmol/L。不同浓度(0.2μmol/L、0.4μmol/L、0.8μmol/L)DSF联合Cu作用于细胞24h后,与Con组相比,DSF/Cu组细胞凋亡率和Bax蛋白表达量明显升高,Bcl-2蛋白表达量降低(均P<0.05);而Cu组与Con组比较差异无统计学意义(P>0.05)。结论:DSF/Cu能抑制人ACCH295R细胞增殖并诱导其凋亡,其机制可能与Bcl-2和Bax表达调控的线粒体凋亡相关通路有关。Objective: To investigate the effects of disulfiram (DSF) combined with copper (Cu) on the proliferation and apoptosis of human adrenocortical carcinoma H295R cells. Methods: H295R cells were divided into blank control group (Con group),Cu group (0.5 μmol/L Cu^2+),and different concentrations of DSF combined with Cu group (DSF/Cu group).The proliferation of H295R cells was detected by MTT assay.The cell apoptosis was analyzed by flow cytometry.The protein expressions of Bcl-2 and Bax were determined by western blotting. Results: The growth inhibition rate of cells was increased in DSF/Cu group in time- and dose-dependent manner,as compared to the blank control group ( P <0.05).The 50% inhibitory concentration (IC 50 ) of H295R cells after intervention for 24 hours was 0.27 μmol/L.After treatment of 0.2 μmol/L,0.4 μmol/L or 0.8 μmol/L DSF and 0.5 μmol/L Cu^2+ for 24 hours,the apoptosis rate of H295R cells and the Bax protein level were increased compared with Con group,while the expression level of Bcl-2 was decreased ( P <0.05).No significant difference was found between the Cu group and the blank control group ( P >0.05). Conclusion: DSF/Cu could inhibit the proliferation of H295R cells and induce apoptosis,and the mechanism might be related to the mitochondrial apoptotic pathway regulated by Bcl-2 and Bax.

关 键 词：双硫仑 铜 肾上腺皮质癌 增殖抑制 细胞凋亡 

分 类 号：R473.73[医药卫生—护理学]