In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402 were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.
XUE Zhi-Gang, LI Jian, YIN Biao, ZHANG Ya-Kun, LIU Xiong-Hao, PAN Qian, LONG Zhi-Gao, DAI He-Ping, XIA Kun, WU Ling-Qian, LIANG De-Sheng, XIA Jia-Hui. p53 Anti-tumor Research in Bel-7402 by Using Human-derived Vector[J]. Progress in Biochemistry and Biophysics,2007,34(5):465-470
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