title: [Recombinant expression of hepatoma associated gene and its protein function].
abstract: OBJECTIVE
  To recombinant express hepatoma associated gene(HTA) and pre-test the function of HTA to determine the role of HTA in the development of liver cancer.
  
  
  METHODS
  HTA338-616 was amplified from HepG2 cells and cloned into the prokaryotic expression vector pET21a(+)-MBP. The proteins MBP and MBP-HTA were induced, purified by His-tag magnetic bead purification kit and identified by Western blot and ELISA. HepG2 cells were stimulated with MBP or MBP-HTA proteins. MTT assay and colony formation assay were employed to examine the proliferation of these cells and the changes of cell cycle distribution were determined by flow cytometry.
  
  
  RESULTS
  The prokaryotic expression plasmid pET21a(+)-MBP-HTA was successfully constructed. We got a 52 kD purified purpose protein.The proliferation of HepG2 cells stimulated with MBP-HTA was significantly higher than those stimulated with MBP and negative controls. HepG2 cells stimulated with MBP-HTA showed significant decrease fraction in G1 phase and increase fraction in S phase, and the cell proliferation was enhanced.
  
  
  CONCLUSIONS
  HTA protein can significantly promote the proliferation of HepG2 cells, which may be related to the promotion of G1 phase to S phase.
authors.affiliation: Cancer Research Institute, Central South University, Changsha 410078, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Song
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Liu
authors.forename: Yalin
authors.initials: Y
authors.lastname: Li
authors.forename: Guancheng
authors.initials: G
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D049109:Cell Proliferation / Q000187:drug effects*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D052138:Genes, Neoplasm / Q000502:physiology*; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006801:Humans; D058619:Maltose-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D016366:Open Reading Frames / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
chemical_list: D058619:Maltose-Binding Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins