肝细胞核因子4α抑制人肝癌细胞株血管内皮生长因子表达和人脐静脉内皮细胞小管形成

Hepatocyte Nuclear Factor 4α Inhibits Expression of Vascular Endothelial Growth Factor in Human Hepatocellular Carcinoma Cell Lines and Tube Formation of Human Umbilical Vein Endothelial Cell

背景:肝细胞核因子4α(HNF4α)在肝脏的发育过程中发挥重要作用。研究证实HNF4α与肝细胞癌(HCC)的发生相关。然而HNF4α对人肝癌细胞株血管内皮生长因子(VEGF)表达和人脐静脉内皮细胞(HUVEC)小管形成的调控作用尚不明确。目的:探讨HNF4α对人肝癌细胞株VEGF表达和HUVEC小管形成的影响。方法:构建过表达HNF4α的慢病毒,并转染人肝癌细胞株HepG 2和SMMC-7721(实验组),以转染慢病毒空载体和未转染的细胞分别作为阴性对照组和空白对照组。分别以qRT-PCR、蛋白质印迹法检测HNF4α、VEGF mRNA和蛋白表达。将HUVEC与HepG2和SMMC-7721细胞不含血清的条件培养基共培养,检测小管形成数量。结果:成功构建了过表达HNF4α的HepG2和SMMC-7721稳转株。与阴性对照组和空白对照组相比,实验组HepG 2和SMMC-7721细胞中VEGF mRNA和蛋白表达均明显降低(P<0.05),HUVEC小管形成数量明显减少(P<0.05)。结论:HNF4α能明显抑制人肝癌细胞株中VEGF表达以及HUVEC小管的形成。

Hepatocyte nuclear factor 4α （HNF4α） plays an important role in the development of liver, and studies demonstrate that it is correXated with the pathogenesis of hepatocellular carcinoma （HCC）. However, the regulatory effect of HNF4α on expression of vascular endothelial growth factor （VEGF） in human HCC cell lines and tube formation of human umbilical vein endothelial cell （HUVEC） is not yet clear. Aims ： To investigate the effect of HNF4α on expression of VEGF in human HCC cell lines and tube formation of HUVEC. Methods： Lentiviral vector overexpressed HNF4α was constructed, and then transfected into HepG2 and SMMC-7721 cells （experimental group）, cells transfected with lentiviral blank vector and cells without transfection were served as negative control group and blank control group, respectively. The mRNA and protein expressions of HNF4ct, VEGF were detected by qRT-PCR and Western blotting, respectively. The conditioned media of HepG2 and SMMC-7721 cells were co-cultured with HUVEC, and number of HUVEC tube formation was measured. Results： HepG2 and SMMC-7721 cells with stable overexpression of HNF4α were successfully established. Compared with negative control group and blank control group, mRNA and protein expressions of VEGF in experimental group were significantly decreased （ P 〈 0.05 ） , and number of HUVEC tube formation was significantly decreased （ P 〈 0.05）. Conclusions ： HNF4α can significantly inhibit the expression of VEGF in HepG2 and SMMC-7721 cells and tube formation of HUVEC.