| 闵雪洁,文 君,赵 丽,刘建军,黄 钢,赵小平.沉默HDGF基因抑制HepG2细胞的增殖和脂质代谢[J].,2018,(11):2006-2011 |
| 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 |
| 投稿时间：2018-01-15 修订日期：2018-01-31 |
| DOI：10.13241/j.cnki.pmb.2018.11.002 |
| 中文关键词: 肝癌衍生生长因子 脂质代谢 增殖 HepG2细胞 |
| 英文关键词: HDGF Lipid Metabolism Proliferation HepG2 cells |
| 基金项目:国家自然科学基金面上项目（81372195） |
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| 中文摘要: |
| 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siRNA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（siRNA-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。 |
| 英文摘要: |
| ABSTRACT 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 siRNA. 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 siRNA-HDGF (or siRNA-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 siRNA-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 siRNA-HDGF. The target lipogenic factors fatty acid synthase (FASN), 3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), stearoyl-CoA desaturase (SCD), and ATP citrate lyase (ACLY) were downregulated to different degrees（P<0.001, P<0.001, P<0.001, P<0.01）. Conclusion: HDGF gene silencing dramatically inhibited the lipid metabolism level and the proliferation of HepG2 cells. |
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