title: Prohibitin is overexpressed in Huh-7-HCV and Huh-7.5-HCV cells harboring in vitro transcribed full-length hepatitis C virus RNA.
abstract: BACKGROUND
  Currently, up-regulated proteins and apoptosis in hepatitis C is a hot topic in exploring the pathogenic mechanism of Heptitis C Virus(HCV). Some recent studies shows that prohibitin is overexpressed in cells expressing HCV core proteins, and up-regulated prohibitin is also found in human hepatoma cell line HCC-M, lung cancer, prostate cancer, and other cancers. Prohibitin is an important member of the membrane protein superfamily, and it plays a role of molecular chaperones in mitochondrial protein stability. Meanwhile, it has a permissive action on tumor growth or acts as an oncosuppressor. Based on our previously established the in vitro HCV cell-culture system (HCVcc), here we aimed to investigate the different expression profiles of prohibitin in Huh-7-HCV and Huh-7.5-HCV cells
  
  
  METHODS
  The total cellular RNA of Huh-7, Huh-7.5, Huh-7-HCV and Huh-7.5-HCV cells were extracted, and then the first-strand cDNA was reversely transcribed. The expression of prohibitin at the mRNA level was assessed by real-time PCR with GAPDH as the control. Furthermore, the expression of prohibitin at the protein level was evaluated by western blot with GAPDH as an internal control.
  
  
  RESULTS
  Our results of real-time PCR showed that the mRNA expression level of prohibitin in Huh-7-HCV cells was 2.09 times higher than that in Huh-7 cells, while, the mRNA level of prohibitin in Huh-7.5-HCV cells was 2.25 times higher than that in Huh-7.5 cells. The results of western blot showed that the protein expression level of prohibitin in Huh-7-HCV cells was 2.38 times higher than that in Huh-7 cells, while the protein expression of prohibitin in Huh-7.5-HCV cells was 2.29 times higher than that in Huh-7.5 cells.
  
  
  CONCLUSIONS
  The expression of prohibitin was relatively high in Huh-7-HCV and Huh-7.5-HCV cells harboring in vitro transcribed full-length HCV RNA.
authors: Department of Infectious Diseases, the Second Affiliated Hospital of Medical School of Xi'an Jiao Tong University, Xi'an, Shannxi 710004, P.R. China. dang212@126.com
authors: Shuang-Suo
authors: SS
authors: Dang
authors: Ming-Zhu
authors: MZ
authors: Sun
authors: E
authors: E
authors: Yang
authors: Meng
authors: M
authors: Xun
authors: Li
authors: L
authors: Ma
authors: Zhan-Sheng
authors: ZS
authors: Jia
authors: Wen-Jun
authors: WJ
authors: Wang
authors: Xiao-Li
authors: XL
authors: Jia
mesh_terms: D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D006526:Hepatitis C / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D010957:Plasmids; D000091402:Prohibitins; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D014779:Virus Replication / Q000235:genetics*
chemical_list: D000091402:Prohibitins; D012333:RNA, Messenger; D012367:RNA, Viral; D012097:Repressor Proteins