PNPTK fusion suicide gene on hepatocellular carcinoma cell killing in rats.doc

 PAGE \* MERGEFORMAT 15 PNPTK fusion suicide gene on hepatocellular carcinoma cell killing in rats Of: Zhoujun Li, Cai Xiaokun Yang Yi Prohibition De-Quan Yao Zhenjiang [Abstract] Objective To analyze the PNP TK fusion suicide gene on HepG2 liver cancer cell killing and its effect on liver cancer cell killing mechanisms. Methods Preparation of recombinant PCR mutagenesis PNP TK fusion gene was inserted into the eukaryotic expression vector pcDNA3 .0 to construct the fusion gene expression vector pcDNA3.0/PNP TK, by restriction analysis, PCR and DNA sequencing of recombinant, G418 selection to obtain stable transfected cell clones resistant pcDNA3.0/PNP TK. RT PCR and Western Blotting detection of PNP TK gene expression in HepG2 cells. trypan blue exclusion method cell growth curve, MTT assay cell sensitivity and corresponding prodrug, respectively, and two prodrugs in a resulting effect of bystander effect. Results PNP TK fusion gene inserted into the pcDNA3.0 vector correctly, pcDNA3.0/PNP TK in HepG2 liver cancer cell lines to achieve the expression. cell clones is very sensitive to the corresponding prodrug. In the two previous Under the combined effects of the drug, pcDNA3.0/PNP TK hepatoma cells a good bystander effect. Conclusion of double suicide gene function with an expression vector pcDNA3.0/PNP TK HepG2 hepatoma cells have a good killing effect expected to be used in liver cancer treatment in vivo. [Keywords:] PNP TK fusion gene; carcinoma; gene therapy Abstract: Objective To investigate the cytotoxic effects of a PNP TK chimeric gene on the basis of PNP suicide gene on HepG2 hepatoma cells and explore the potential mechanism of their killing effects.Methods A fusion suicide gene, PNP TK, was produced by site directed mutagenesis technique. And then it was inserted into pcDNA3.0 to construct a vector harboring a chimeric gene, pcDNA3.0/PNP TK. The recombinant was identified by recombinant enzyme, PCR and sequencing. Then it was transfecte