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AIM: To investigate the effects of p16 gene on biologicalbehavious in hepatocellular carcinoma cells.METHODS: HCC cell lines SNU-449 and HepG2.2.15 wereinfected respectively by a replication defective, recombinantretrovirus capable of producing a high level of p16 proteinexpression (pCLXSN-p16). G418 resistant stable P16 proteinexpression cell lines were selected. And the biological behavioursof the p16 gene transfected HCC cells were observed.RESULTS: Initialin vitro experiments in HCC cell line SNU-449 with loss of p16 protein expression demonstrated thepCLXSN-p16 treatment significantly inhibited cell growth.But there was no treatment effect when the pCLXSN-p16was used in another HCC cell line HepG2.2.15 which haspositive p16 protein expression. Subsequent study in a nudemouse model demonstrated that the p16 gene transfectedSNU-449 had a lower succeeding rate in the first timeestablishment of tumors and grew more slowly in the nudemice when compared with non-transfected SNU-449.Moreover, the nude mice inoculated with transfected SNU-449 had a longer surviving time than those inoculated withnon-transfected SNU-449.CONCLUSION: Our results show that the p16INK4a genetransfer can inhibit the proliferation and reduce the invasionability of hepatocellular carcinoma.