MEG3和腺苷对HepG2细胞自噬和增殖的影响  

Effects of MEG3 and adenosine on autophagy and proliferation of HepG2 cells

作　　者：蒲泽锦[1] 李国平 詹浩炼 周小涛[1] 郭益添[1] 项梦琦 刘丽璇 谭辉[2] 吴灵飞[1] PU Ze-jin;LI Guo-ping;ZHAN Hao-lian;ZHOU Xiao-tao;GUO Yi-tian;XIANG Meng-qi;LIU Li-xuan;TAN Hui;WU Ling-fei(Department of Gastroenterology,The Second Affiliated Hospital of Shantou University Medical College,Shantou 515041,China;Department of Radiology,The Second Affiliated Hospital of Shantou University Medical College,Shantou 515041,China)

机构地区：[1]汕头大学医学院第二附属医院消化内科,广东汕头515041 [2]汕头大学医学院第二附属医院放射科,广东汕头515041

出　　处：《中国病理生理杂志》2018年第9期1706-1710,共5页Chinese Journal of Pathophysiology

基　　金：广东省科技计划(No.2014A020212284);广东省自然科学基金资助项目(No.2014A030313470)

摘　　要：目的:研究母系表达基因3(MEG3)过表达和腺苷对人肝细胞癌HepG2细胞自噬和增殖的影响,并探讨MEG3和腺苷影响HepG2细胞自噬的可能机制和抑制细胞增殖的效果。方法:常规培养HepG2细胞,分为对照组、MEG3组(MEG3慢病毒转染)、腺苷组(1 mmol/L腺苷处理)和MEG3+腺苷组(腺苷组中加入MEG3慢病毒转染)。Western blot检测LC3-Ⅱ、LC3-Ⅰ和哺乳动物雷帕霉素靶蛋白(mTOR)等蛋白的表达,MDC染色观察自噬体数量,CCK-8法观察细胞的活力,细胞计数仪检测活细胞数量。结果:与对照组比较,MEG3组和腺苷组中HepG2细胞内LC3-Ⅱ减少,LC3-Ⅱ/LC3-Ⅰ比值降低,mTOR表达增加,HepG2细胞的活力降低(P<0.05),自噬体减少。与MEG3组和腺苷组比较,MEG3+腺苷组中HepG2细胞内LC3-Ⅱ和LC3-Ⅱ/LC3-Ⅰ比值进一步降低,mTOR表达进一步增加,HepG2细胞的活力进一步降低,自噬体减少更加显著(P<0.05或P<0.01)。MEG3组和腺苷组中HepG2活细胞数在24～72 h较对照组均明显降低(P<0.01),在MEG3+腺苷组中降低更为明显(P<0.01)。结论:MEG3过表达和低浓度腺苷可激活mTOR通路,抑制HepG2细胞自噬和增殖。MEG3增强了腺苷对HepG2细胞的作用。AIM:To study the effects of maternal expressed gene 3(MEG3)and adenosine on the autophagy and proliferation of human hepatocellular carcinoma HepG2 cells,and to explore the possible mechanisms of autophagy and the effect on the proliferation of HepG2 cells induced by MEG3 and adenosine.METHODS:HepG2 cells were cultured according to the conventional cultural method,and divided into control group,MEG3 group(the cells were transfected with MEG3 lentivirus),adenosine group(the cells were treated with 1 mmol/L adenosine)and MEG3+adenosine group(the cells were treated with 1 mmol/L adenosine and MEG3 lentivirus).The protein expression of LC3-Ⅱ,LC3-Ⅰand mammalian target of rapamycin(mTOR)was determined by Western blot.MDC staining was used to observe the number of autophagosomes.The cell viability was measured by CCK-8 assay,and the number of viable cells were counted by automated cell counter.RESULTS:Compared with control group,LC3-Ⅱexpression and the ratio of LC3-Ⅱ/LC3-Ⅰwere decreased,mTOR expression was increased(P<0.05),the viability of HepG2 cells was decreased,and the number of autophagosomes were reduced in MEG3 group and adenosine group.In MEG3+adenosine group,LC3-Ⅱexpression and the ratio of LC3-Ⅱ/LC3-Ⅰwere decreased significantly(P<0.01),mTOR expression was increased significantly(P<0.01),and the viability and autophagosomes of HepG2 cells were reduced markedly as compared with MEG3 group and adenosine group.After treated with MEG3 and adenosine for 24~72 h,the viable HepG2 cells reduced significantly in MEG3 group and adenosine group(P<0.01),especially in MEG3+adenosine group(P<0.01).CONCLUSION:MEG3 overexpression and low concentration of adenosine activate the mTOR pathway,and inhibit the autophagy and proli-feration of HepG2 cells.MEG3 enhances the effect of adenosine on HepG2 cells.

关 键 词：母系表达基因3 腺苷 自噬 哺乳动物雷帕霉素靶蛋白 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤] R363[医药卫生—临床医学]