上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

miR-143通过靶向FASN抑制肝癌HepG2细胞脂质形成

李卫萍1，李锦平1，李元宏2，任来峰2，宋维芳3   

  1. 山西医科大学 汾阳学院 1.药理教研室, 2.生物化学与分子生物学教研室， 3.病理生理教研室， 汾阳 032200
  • 出版日期:2015-03-28 发布日期:2015-03-26
  • 作者简介:李卫萍(1978—)， 女， 讲师， 硕士生; 电子信箱: liweiping0929@163.com。
  • 基金资助:

    中国肝炎防治基金(TQGB20120013)

Inhibition of the lipogenesis of hepatocellular carcinoma HepG2 cells by miR-143 via suppressing FASN

LI Wei-ping1, LI Jin-ping1, LI Yuan-hong2, REN Lai-feng2, SONG Wei-fang3   

  1. 1.Department of Pharmacology, 2.Department of Biochemistry and Molecular Biology, 3.Department of Pathophysiology, Shanxi Medical University Fenyang College, Fenyang 032200, China
  • Online:2015-03-28 Published:2015-03-26
  • Supported by:

    Hepatitis Prevention and Control Foundation of China, TQGB20120013

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摘要：

目的 研究miR-143对肝癌HepG2细胞脂质形成的影响及其分子机制。方法 应用qRT-PCR方法检测miR-143在临床肝癌和癌旁组织以及肝癌细胞系和正常肝细胞系中的表达。通过油红O染色实验检测miR-143对肝癌细胞脂质形成的影响。Western blotting检测miR-143对肝癌细胞中脂肪酸合成酶(FASN)表达的影响。通过报告基因实验检测miR-143及其抑制子对报告基因载体FASN-CDS活性的影响。结果 qRT-PCR结果表明在临床肝癌组织和HepG2细胞中miR-143的表达显著低于癌旁组织和Chang liver肝细胞系（P<0.01）。油红O染色实验结果显示，miR-143能显著降低肝癌细胞中脂质的形成。qRT-PCR和Western blotting结果证实，miR-143可以显著下降FASN的mRNA和蛋白表达水平。报告基因检测结果表明，在HepG2细胞中miR-143可以显著降低FASN-CDS报告基因的活性（P<0.01）；相反，miR-143抑制子可以显著升高FASN-CDS的活性（P<0.01）。在HepG2细胞中miR-143抑制子可以上调FASN的表达并促进脂质形成。结论 miR-143可以通过抑制FASN的表达阻碍肝癌细胞脂质形成。

关键词: miR-143, 肝癌, 脂质形成, FASN

Abstract:

Objective To investigate the effects of miR-143 on the lipogenesis of hepatocellular carcinoma HepG2 cells and the molecular mechanism. Methods Expressions of miR-143 of the clinical hepatocarcinoma tissue, para cancer tissue, hepatocellular carcinoma cell line, and normal liver cell line were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The effect of miR-143 on the lipogenesis of hepatocellular carcinoma cells was detected by the oil red O staining assay. The effect of miR-143 on the expression of fatty acid synthase (FASN) of hepatocellular carcinoma cells was detected by the Western blotting. The effects of miR-143 and its inhibitor on the activity of reporter gene vector FASN-CDS were detected by the reporter gene assays. Results The results of qRT-PCR showed that the expression of miR-143 of the clinical hepatocarcinoma tissue and HepG2 cells significantly lower than that of the para cancer tissue and Chang liver cell line (P<0.01). The results of oil red O staining assay indicated that miR-143 significantly decreased the lipid formation of hepatocellular carcinoma cells. The results of qRT-PCR and Western blotting showed that miR-143 significantly decreased mRNA and protein levels of FASN. The results of reporter gene assays indicated that miR-143 in HepG2 cells significantly decreased the activity of FASN-CDS (P<0.01), while the inhibitor of miR-143 significantly increased the activity of FASN-CDS (P<0.01). The inhibitor of miR-143 in HepG2 cells was able to up-regulate the expression of FASN and enhance the lipogenesis. Conclusion MiR-143 can inhibit the lipogenesis of hepatocellular carcinoma cells by suppressing the expression of FASN.

Key words: miR-143, hepatoma, lipogenesis, FASN