title: [The expression vector of murine secreting IL-1β  promotes proliferation and migration of Hepa1-6 hepatoma cells].
abstract: OBJECTIVE
  To establish a murine secreted mature peptide IL-1β expression vector, transfect into Hepa1-6 hepatoma cells, and analyze the effect of recombinant IL-1β on proliferation, migration, and its specific expression in Hepa1-6 hepatoma cells.
  
  
  METHODS
  The murine AFP signal peptide encoding sequence and mature IL-1β encoding fragment were linked together through overlapping PCR, and the chimeric DNA sequence was then inserted into a liver specific expression vector pLIVE(TM); to make a recombinant pLIVE-smIL-1β which expressed secreted murine IL-1β of classical pathway. pLIVE-smIL-1β, pLIVE(TM); and pLIVE-lacZ were transfected into Hepa1-6 by jetPEI respectively. Transfection of the vectors were detected by β-gal staining using pLIVE-lacZ transfectants. Cells treated with 5 μg/mL lipopolysaccharide were used as positive control and 3 μmol/L monesin was added into culture system to block classical pathway secretion, then sandwich ELISA was employed to detect the IL-1β levels both in supernatant and in cytoplasm of each group of transfected cells. The proliferation of Hepa1-6 hepatoma cells was determined by MTT assay and migration of Hepa1-6 cells was assessed by scratch test in vitro.
  
  
  RESULTS
  pLIVE-smIL-1β vector successfully expressed murine IL-1β in Hepa1-6 cells. Expression of the recombinant vector peaked at day 3 as indicated in a β-gal staining method. After transfection, compared with Hepa1-6/mock cells, IL-1β expression levels both in supernatant and in cytoplasm of Hepa1-6/smIL-1β cell were significantly increased detected by ELISA. The proliferation of Hepa1-6/smIL-1β group was markedly promoted in vitro detected by MTT assay.
  
  
  CONCLUSIONS
  The recombinant expression vector can secret IL-1β through classical pathway which significantly promoted proliferation and migration of hepatoma cells in vitro.
authors.affiliation: Key Laboratory of Molecular Immunology, Weifang Medical College, Weifang, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Tian
authors.forename: Zhi-juan
authors.initials: ZJ
authors.lastname: Lin
authors.forename: Ping
authors.initials: P
authors.lastname: Sun
authors.forename: Yan-fei
authors.initials: YF
authors.lastname: Liu
authors.forename: Ji-yu
authors.initials: JY
authors.lastname: Ju
authors.forename: Shu-juan
authors.initials: SJ
authors.lastname: Liang
mesh_terms: D000818:Animals; D002465:Cell Movement; D049109:Cell Proliferation; D005822:Genetic Vectors; D053583:Interleukin-1beta / Q000235:genetics* / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D051379:Mice
chemical_list: D053583:Interleukin-1beta