The Effect And Mechanism Of Exogenous P16 Expression On Biological Behavior Of Human Hepatocellular Carcinoma

Objective: To construct human pl6 gene eukaryotic expression vector.To investigate the effect of exogenous pl6 expression on biological behavior of human hepatocellular carcinoma and its mechanism.Methods: pcDNA3. 1(+)-pl6 eukaryotic expression plasmid was constructedand transfected into SMMC-7721 cells which retains down- regulated pl6 gene expression. Integration and expression of exogenous pl6 gene were confirmed by RT-PCR and immunocytochemistry. The cell county, flow cytometry , immunocytochemistry and western blot were used to observe the biological behaviors of pl6 gene transfected hepatocellular carcinoma cells and to investigate the effect of exogenous pl6 expression on CyclinDl and pRb in SMMC-7721 cells.Results: The pcDNA3. 1(+)-pl6 eukaryotic expression plasmid was obtainedsuccessfully . The cloned pl6cDNA coding sequence was shown to be consistent with reported sequence derived from Genbank. The growth of the cells transfected with pl6 was decreased and the percentage of the cell in G0/G1 phase was increased. The expression of cylinDl and phosphorylated pRb were down-regulated in SMMC-7721 cells which transfected with pl6 gene.Conclusion:1 , The pcDNA3. 1(+)-p16 eukaryotic expression plasmid was obtainedsuccessfully .2, Exogenous pl6 gene could be expressed in SMMC-7721. It may arrest cellcycle in G0/G1 phase and suppress cell growth.3, The mechanism may be that pl6 gene down-regulate the expression of...