摘要
目的观察转录激活因子4(ATF4)过表达对人肝癌HepG2细胞凋亡的影响,并探讨其机制。方法构建ATF4表达质粒,采用脂质体瞬时转染HepG2细胞。实验分为空白对照组、空质粒组和ATF4质粒组。采用流式细胞术检测细胞凋亡率,检测HepG2细胞培养液中乳酸含量,Western blot检测Warburg效应蛋白M2型丙酮酸激酶(PKM2)的表达。结果与空质粒组比较,ATF4质粒组HepG2细胞凋亡率显著增加,细胞培养液中乳酸含量显著降低,HepG2细胞中PKM2蛋白的表达显著下调(均P<0.05)。结论过表达ATF4促进HepG2细胞的凋亡,其机制可能与抑制Warburg效应有关。
Objective To observe the effect of overexpression of activating transcription factor 4（ATF4） on the apoptosis of HepG2 cells and explore the mechanism. Methods ATF4 expression plasmid was built. Hepatocellular carcinoma line HepG2 cells were transfected with ATF4 expression plasmid through liposome. The plasmids were divided into blank control group, empty plasmid group and ATF4 plasmid group. The apoptosis rate was detected by flow cytometry.The content of lactate in cell culture medium was tested. The expression of Warburg effect protein pyruvate kinase subtype M2（PKM2） was measured by Western blot. Results Compared with the empty plasmid group, the apoptosis rate of HepG2 cells increased significantly, the content of lactate in cell culture medium decreased significantly and the expression of PKM2 decreased significantly（all P〈0.05）. Conclusion Overexpression of ATF4 promotes the apoptosis of hepatocellular carcinoma line HepG2 cells, the mechanism may be related to inhibition of Warburg effect.
作者
肖婷焱
段无暇
许紫薇
周寿红
曾斌
XIAO Tingyan;DUAN Wuxia;XU Ziwei;ZHOU Shouhong;ZENG Bin(Department of Gastroenterology,the First Affiliated Hospital of University of South China,Hu＇nan Province,Hengyang 421001,China;Teaching and Research Office of Physiology,Medical College,University of South China,Hu＇nan Province,Hengyang 421001,China)
出处
《中国医药导报》
CAS
2018年第19期12-15,29,共5页
China Medical Herald
基金
湖南省自然科学基金资助项目(08JJ3032)
湖南省卫生厅科研计划项目(B2007100)
