title: HCV core protein promotes hepatocyte proliferation and chemoresistance by inhibiting NR4A1.
abstract: This study investigated the effect of HCV core protein on the proliferation of hepatocytes and hepatocellular carcinoma cells (HCC), the influence of HCV core protein on HCC apoptosis induced by the chemotherapeutic agent cisplatin, and the mechanism through which HCV core protein acts as a potential oncoprotein in HCV-related HCC by measuring the levels of NR4A1 and Runt-related transcription factor 3 (RUNX3), which are associated with tumor suppression and chemotherapy resistance. In the present study, PcDNA3.1-core and RUNX3 siRNA were transfected into LO2 and HepG2 cells using Lipofectamine 2000. LO2-core, HepG2-core, LO2-RUNX3 (low) and control cells were treated with different concentrations of cisplatin for 72 h, and cell proliferation and apoptosis were assayed using the CellTiter 96(®)Aqueous Non-Radioactive Cell Proliferation Assay Kit. Western blot and real time PCR analyses were used to detect NR4A1, RUNX3, smad7, Cyclin D1 and BAX. Confocal microscopy was used to determine the levels of NR4A1 in HepG2 and HepG2-core cells. The growth rate of HepG2-core cells was considerably greater than that of HepG2 cells. HCV core protein increased the expression of cyclin D1 and decreased the expressions of NR4A1 and RUNX3. In LO2 - RUNX3 (low), the rate of cell proliferation and the level of cisplatin resistance were the same as in the LO2 -core. These results suggest that HCV core protein decreases the sensitivity of hepatocytes to cisplatin by inhibiting the expression of NR4A1 and promoting the expression of smad7, which negatively regulates the TGF-β pathway. This effect results in down regulation of RUNX3, a target of the TGF-β pathway. Taken together, these findings indicate that in hepatocytes, HCV core protein increases drug resistance and inhibits cell apoptosis by inhibiting the expressions of NR4A1 and RUNX3.
authors.affiliation: Department of Oncology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China. Electronic address: yongshengtanwhu@126.com.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Tan
authors.affiliation: Department of Oncology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China. Electronic address: liyansd2@163.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000473:pathology / Q000821:virology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D050677:Core Binding Factor Alpha 3 Subunit / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019938:Cyclin D1 / Q000378:metabolism; D024882:Drug Resistance, Viral; D056945:Hep G2 Cells; D016174:Hepacivirus / Q000472:pathogenicity* / Q000502:physiology*; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology* / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000821:virology; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D051906:Smad7 Protein / Q000378:metabolism; D014758:Viral Core Proteins / Q000502:physiology*
chemical_list: D050677:Core Binding Factor Alpha 3 Subunit; C539821:NR4A1 protein, human; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; C493729:Runx3 protein, human; C497806:SMAD7 protein, human; D051906:Smad7 Protein; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D019938:Cyclin D1; D002945:Cisplatin
keywords: Chemoresistance; HCV core protein; Hepatocellular carcinoma; NR4A1; Oncogene; RUNX3