title: [Effect of glypican-3 on the proliferation of human hepatoma cell line MHCC97-L in vitro].
abstract: OBJECTIVE
  To construct glypican-3 (GPC3)-green fluorescent protein eukaryotic expression vector pEGFP-c3-GPC3, and analyze the effect of GPC3 on the proliferation of human hepatoma cell line MHCC-97L.
  
  
  METHODS
  The eukaryotic expression vector pEGFP-c3-GPC3 was constructed with recombinant DNA technique and transfected into MHCC-97L cells via Lipofectamine 2000. The cells stably expressing GPC3 were screened by flow cytometry and G418. The mRNA expression of GPC3 was detected by RT-QPCR method, and the protein expression by Western blotting and fluorescence microscope. The effect of GPC3 gene on the growth of the cells was examined by MTT assay.
  
  
  RESULTS
  Restriction endonuclease analysis and DNA sequencing verified correct construction of the recombinant plasmid. The green fluorescence was detected in the transfected MHCC-97L cells under fluorescence microscope. RT-QPCR and Western blotting both confirmed successful expression of GPC3 in MHCC-97L cells. The growth curve showed a significant acceleration of the proliferation of the transfected MHCC97-Lsol;GPC3 cells as compared with MHCC97-L and MHCC97-L/C3 cells (P<0.001).
  
  
  CONCLUSIONS
  We have successfully constructed the eukaryotic expression vector pEGFR-c3-GPC3, which allows stable GPC3 expression in MHCC97-L/GPC3 cells. The upregulation of GPC3 expression can stimulate the growth of hepatoma cell line MHCC97-L in vitro.
authors.affiliation: Oncology Center, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. qruzhai1@126.com
authors.forename: Ru-zhai
authors.initials: RZ
authors.lastname: QIN
authors.forename: Fei-ye
authors.initials: FY
authors.lastname: LIU
authors.forename: Bin
authors.initials: B
authors.lastname: CHEN
authors.forename: Jian-ming
authors.initials: JM
authors.lastname: XIE
authors.forename: Yang
authors.initials: Y
authors.lastname: YANG
authors.forename: Da-yong
authors.initials: DY
authors.lastname: ZHEGN
authors.forename: Rong-cheng
authors.initials: RC
authors.lastname: LUO
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D005822:Genetic Vectors; D053673:Glypicans / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D010957:Plasmids; D014162:Transfection
chemical_list: D053673:Glypicans; D049452:Green Fluorescent Proteins