奥沙利铂诱导的人肝癌细胞株HepG2自噬潮增加及其对细胞生存率的影响  

The enhancement of autophagic flux induced by oxaliplatin in hepatocarcinoma cell line HepG2 and its influence on cell proliferation

作　　者：李媛媛[1] 马泰[1] 仲飞[1] 范璐璐[1] 孙国平[1] 

机构地区：[1]安徽医科大学第一附属医院肿瘤内科,安徽合肥230032

出　　处：《肿瘤》2013年第2期132-137,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:81071986);国家自然科学基金资助项目(编号:81272739);教育部高等学校博士学科点专项科研基金--博导类资助项目(编号:20113420110002);安徽高校省级自然科学研究重点资助项目(编号:KJ2011A154)

摘　　要：目的:研究奥沙利铂(oxaliplatin,Oxa)对人肝癌HepG2细胞自噬功能的影响,并初步探讨自噬在Oxa诱导细胞死亡中的作用。方法:体外常规培养的人肝癌细胞HepG2,分别以Oxa单药和Oxa+自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)或氯喹(chloroquine,CQ)联合作用后,采用CCK-8(cell counting kit-8)法检测细胞活力的变化;透射电子显微镜下观察Oxa作用后HepG2细胞内自噬体的形成;蛋白印迹法检测自噬标志蛋白LC3-Ⅰ向LC3-Ⅱ的转变。结果:Oxa可引起HepG2细胞死亡并呈现出剂量依赖关系,当与3-MA或CQ联用时均能对HepG2细胞的生长产生明显的抑制作用;Oxa作用后可诱导HepG2细胞内自噬小体的形成,并出现自噬相关蛋白LC3-Ⅱ表达的增多,CQ抑制自噬降解通路后LC3-Ⅱ蛋白则明显增加(自噬潮增加);3-MA则可以抑制HepG2细胞自噬体的形成。结论:Oxa可以增强HepG2细胞的自噬功能,诱导自噬体形成增加;不同靶点的自噬抑制剂3-MA或CQ均可明显增加Oxa对肝癌细胞的生长抑制作用,增强Oxa的肿瘤杀伤作用。Objective： To investigate the change in function of autophagy in hepatocarcinoma cell line HepG2 induced by Oxa （oxaliplatin）, and to explore the role of autophagy in Oxa-induced cell death. Methods： The HepG2 cells were routinely cultured in vitro and treated either with different concentrations of Oxa alone or in combination with the autophagy inhibitor 3-MA （3-methyladenine） or CQ （chloroquine）. The cell viability was detected by CCK-8 （cell counting kit-8） assay. The formation of autophagosomes was observed directly under TEM （transmission electron microscope）. Western-blotting was used to track the conversion of autophagic marker proteins LC3-Ⅰ to LC3-Ⅱ. Results： CCK-8 assay demonstrated that Oxa could dose-dependently induce the death of HepG2 cells, and when it was administrated in combination with 3-MA or CQ, Oxa could also significantly inhibit the growth of HepG2 cells. The formation of autophagosomes in HepG2 cells after Oxa treatment could be observed under TEM, as well as an increase in expression of LC3-Ⅱ was found. The expression of LC3-Ⅱ increased markedly after the inhibition of degradation pathway induced by CQ, which reflected an enhancement of autophagic flux induced by Oxa. 3-MA could inhibit the formation of autophagosomes in HepG2 cells. Conclusion： Oxa can enhance the function of autophagy in HepG2 cells and induce an increase in autophagosome formation. The autophagy inhibitors 3-MA and CQ can both facilitate the proliferative inhibition of hepatocarcinoma cell line HepG2 induced by Oxa, and they can also increase the antitumor effect of Oxa.

关 键 词：肝肿瘤 实验性 自噬 奥沙利铂 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤] Q256[医药卫生—临床医学]