P53 Anti-tumor Research In Bel-7402 By Using Human-derived Vector

Hepatic cell carcinoma (HCC) is a tumor with high lethality. Most HCC patients can not survive for more than one year except some at early stage. Conventional treatment can only be effective in some patients with HCC at early stage. Cancer gene therapy, as a new therapeutic theory, attracts many researchers' attention once it appears.Objects: Safe, efficient and specific vector is the key to cancer gene therapy. In this research, we investigated the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector phrn in liver cancer cell Bel-7402.Methods: We constructed the report plasmid pchEGFP and tumor suppressor plasmids pchp53_Arg and pchp53_Pro by inserting the expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53_Arg and CMVe+hTERTp+p53_Pro into phrn respectively. Twenty four hours after cell transfection of those plasmids by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blotting were performed to study the p53 expression pattern. We also detected the cell apoptosis by HOECHST 33258 and ANNEXIN V-FITC/PI staining.Results: The expression of GFP and p53 proteins in Bel-7402 was detected, but apparent cell apoptosis could not be found.Conclusions: The research indicated that the recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might be resulted from the down regulation effect of the wild type p53 on hTERTp...