CONSTRUCTION OF p16 GENE RECOMBINANT EXPRESSION VECTOR AND ITS ANTITUMORIGENIC EFFECTS ON HUMAN GLIOMA CELLS

Objective:To explore the effect of p16 gene on the growth inhibition of glioma cell line SHG-44 .Methods:We cloned a p16cDNA into the pcDNA3 vector at the sites of BamH Ⅰ and Xho Ⅰ to gain a p16 gene recombinant expression vector plasmid . The p16 gene recombinant plasmid was transfected into the glioma cell line ,In these glioma cells ,the p16 gene was homozygously deleted .Results:Expression of exogenous p16 gene in SHG-44 cell was identified by PCR and Western blot analysis .The growth rate of SHG-44 transfected with p16 gene (SHG-44-p16) was markedly suppressed . Colony formation in soft agar was also decreased significantly .Cell cycle analysis by flow cytometry showed that the number of cell in G 1-G 0 phase of SHG-44 cells was significantly increased while cells in S and G 2 M phase was decreased compared to that of control SHG-44 cells .Conclusion:Transfection of wild-type p16 gene into p16 gene-deleted glioma cells can restore its suppressive effect on cell growth by arrest of cell cycle at G 1 phase.[