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Adenovirus E1a gene enhances P16 gene-induced apoptosis of hepatocellular carcinoma SMMC-7721 cells
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Abstract:
Objective :To investigate the synergistic effect of anti-cancer P16 gene and adenovirus E1a gene on apoptosis and proliferation of hepatocellular carcinoma SMMC-7721 cells, and to explore the novel therapeutic strategy for tumor gene therapy. Methods: Eukaryotic expression plasmid pDC315-E1a and adenoviral vector AdCMV-P16 were constructed. The expression of P16 and E1a in SMMC-7721 cells after pDC315-E1a transfection or AdCMV-P16 infection was determined by RT-PCR and immunofluorescent labeling. SMMC-7721 cell transplanted tumors in nude mice was established. The effect of pDC315-E1a and AdCMV-P16 alone or incombination on tumor growth was observed, and the expressions of P16 and E1a in transplanted tumor tissues and apoptosis of transplanted tumor cells were determined by immunohistochemistry and TUNEL assay, respectively. Results: SMMC-7721 cells showed positive expression of both mRNA and protein levels of E1a and P16 after pDC315-E1a transfection or AdCMV-P16 infection, respectively. Compared with the control group, the apoptosis rate of transplanted tumor cells was (14.3±2.5)% (P<0.01) and tumor inhibitory rate was 361% (P<0.01) in AdCMV-P16 therapy group; those in pDC315-E1a therapy group was (8.5±2.9)% (P<0.01) and 17.1% (P>0.05); and in AdCMV-P16 combined pDC315-E1a therapy group was (27.3±6.3)% (P<0.01) and 57.2% (P<0.01), respectively. Conclusion: Adenovirus E1a gene can increase P16-induced apoptosis and cell growth inhibition in SMMC-7721 cell transplanted tumors, and thus enhance the efficacy of P16 gene therapy.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81071866, No.81172303)
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