Original Articles
Effect of hepatocyte nuclear factor 1α on expression profile of hepatocellular carcinoma cells HuH7
Chenhong Ding, Longfei Deng, Jian Wang, Weifen Xie, Xin Zhang
Published 2015-03-15
Cite as Chin J Dig, 2015, 35(3): 193-197. DOI: 10.3760/cma.j.issn.0254-1432.2015.03.013
Abstract
ObjectiveTo investigate the regulation effect of hepatocyte nuclear factor (HNF) 1α on the gene expression profile and the signal pathways in HuH7 cells.
MethodsThe expression of HNF1α was increased or decreased in HuH7 cells by Lenti-virus carrying HNF1α or shHNF1α. The expression profile of the cells after treated was examined by microarray technology. The difference expressed gene regulated by HNF1α were screened and the pathway was analyzed with DAVID software and related analysis system. The regulation effect of HNF1α on transforming growth factor (TGF)β signal pathway was detected by reporter gene test and the regulation role of HNF1α on related genes of TGFβ signal pathway was determined by real-time polymerase chain reaction (PCR) and Western blotting assay.
ResultsThe expression of HNF1α in HuH7 cells was significantly up-regulated by Lenti-virus carrying HNF1α gene (Lenti-HNF1α) and which was down regulated by Lenti-virus with shHNF1α gene (Lenti-shHNF1α). Expression profile analysis revealed that 339 genes were positively up regulated two times by HNF1α and 325 genes were negatively down regulated two times. Signal pathway analysis revealed that HNF1α regulated drug metabolism, biosynthesis of unsaturated fatty acids and glycolysis/gluconeogenesis metabolism signal pathways. Moreover, it also involved in the regulation of TGFβ、nuclear factor (NF)-κB and p53 tumor-related signal pathways. Furthermore, Luciferase reportor gene experiment indicated that up-regulated HNF1α could inhibit the activation of TGFβ signal pathway. And the results of real-time PCR and Western blotting verified that up-regulated HNF1α could inhibit TGFβ signal pathway related gene c-myc and TGFβ1 and then inhibited the activation of TGFβ signal pathway.
ConclusionHNF1α broadly affects the gene expression profile and the tumor genesis and development related signal pathways in HuH7 cells, furthermore, HNF1α can inhibit the activation of TGFβ signal pathway.
Key words:
Hepatocyte nuclear factor 1-alpha; Gene expression profiling; TGF-β signal pathway
Contributor Information
Chenhong Ding
School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
Longfei Deng
Jian Wang
Weifen Xie
Xin Zhang
