沉默HDGF基因抑制HepG2细胞的增殖和脂质代谢  

Down Regulation of HDGF Suppressed the Proliferation and Lipid Metabolism of HepG2 Cells

作　　者：闵雪洁 文君[1] 赵丽[1] 刘建军[1] 黄钢[1] 赵小平[1] MIN Xue-jie;WEN Jan;ZHAO Li;LIU Jian-jun;HUANG Gang;ZHAO Xiao-ping(Department of Nuclear Medicine,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai,200127,China)

机构地区：[1]上海交通大学医学院附属仁济医院核医学科,上海200127

出　　处：《现代生物医学进展》2018年第11期2006-2011,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81372195)

摘　　要：目的:探讨肝癌衍生生长因子(HDGF)对HepG2细胞增殖和脂质代谢的影响。方法:用脂质体包裹si RNA的方法沉默HDGF基因,用实时荧光定量PCR法和蛋白质免疫印迹法检测HDGF在mRNA和蛋白水平的变化,检测细胞总甘油三酯、胆固醇含量并用油红O染色,CCK-8检测及琼脂糖凝胶克隆形成,实时荧光定量PCR法检测脂质代谢相关酶的mRNA表达。结果:将靶向HDGF小干扰(si RNA-HDGF)转染到HepG2细胞后,可明显抑制HDGF的mRNA表达(P<0.001)和蛋白表达。HDGF蛋白抑制后,细胞增殖在48 h(P<0.01)、72 h(P<0.001)和96 h(P<0.001)均明显降低;细胞内总甘油三酯及胆固醇水平也明显降低(P<0.05,P<0.01)。此外,油红O染色显示细胞内脂滴有明显的减少。脂质代谢相关酶脂肪酸合成酶(FASN)、羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)、硬脂酰辅酶A去饱和酶(SCD)及ATP-柠檬酸裂解酶(ACLY)的mRNA表达均明显降低(P<0.001,P<0.001,P<0.001,P<0.01)。结论:抑制HDGF的表达可明显降低HepG2细胞内脂质代谢水平并抑制其增殖。Objective： To identify the effect of Hepatoma-derived growth factor（HDGF） on the proliferation and lipid metabolism of HepG2 cells. Methods： We silenced HDGF using si RNA. The efficiency of HDGF silencing in mRNA and protein levels were detected by real-time fluorescent quantitative PCR and Western blot, respectively. The total triglycerides and cholesterols levels were detected, and Oil red O staining, CCK-8 assay, soft agar colony formation assay were performed, real-time fluorescent quantitative PCR was used to analyze the mRNA expressions of metabolism related factors. Results： After the si RNA-HDGF（or si RNA-NC as control）was successfully transfected into HepG2 cells, the expression levels of HDGF mRNA（P〈0.001） and protein were significantly downregulated. HepG2 cells transfected with si RNA-HDGF at 48 h（P〈0.01）, 72 h（P〈0.001）and 96 h（P〈0.001）were decreased; total triglycerides and cholesterols levels in intracellular were lower than those of the control group. Furthermore, Oil red O staining showed that the number of intracellular lipids droplets was obviously decreased in HepG2 cells transfected with si RNA-HDGF. The target lipogenic factors fatty acid synthase（FASN）, 3-Hydroxy-3-methylglutaryl-coenzyme A reductase（HMGCR）, stearoyl-Co A desaturase（SCD）, and ATP citrate lyase（ACLY） were downregulated to different degrees（P〈0.001, P〈0.001, P〈0.001, P〈0.01）. Conclusions：HDGF gene silencing dramatically inhibited the lipid metabolism level and the proliferation of HepG2 cells.

关 键 词：肝癌衍生生长因子 脂质代谢 增殖 HEPG2细胞 

分 类 号：R-33[医药卫生] R735.7