重组腺病毒介导的人野生型p53、GM—CSF和B7—1基因在肝癌细胞中的表达

目的：观察腺病毒载体对肝癌细胞的转染效率及其介导的人野生型p53，GM－CSF和B7－1基因在肝癌细胞中的表达。方法：不同MOI的Ad-GFP感染肝癌细胞，48h后计算感染效率；BB－102以50MOI感染肝癌细胞，48h后免疫组化法及western blot检测p53表达，ELISA检测GM0CSF的含量，FACS测定B7－1的表达。结果MOI为50pfu/细胞时对肝癌细胞的转染效率达80％以上，目的基因均可在肝癌细胞中高效表达。结论：腺病毒载体对肝癌细胞具有较高的转染效率，目的基因均可在肝癌细胞中高效表达，为进一步研究BB－102在肝癌治疗中的应用奠定了基础。

Wild-type p53,GM-CSF,B7-1 genes transduced with recombinant adenovirus expression in hepatoma cell lines in vitro

Objective To evaluate the transferring efficiency of recombinant adenoviral vector(BB|102) expressing the human wild type p53,GM|CSF and B7|1 genes into hepatocellular carcinoma(HCC) cell lines in vitro. Methods HCC cell lines were transferred with mock adenoviral vector expressing green fluorescent protein(Ad|GFP) at a series concentrations of MOI. p53, GM|CSF, and B7|1 expressing in HCC cells were determined by western blot, ELISA method and flow|cytometric analyses respectively. Results When 50MOI of recombinant adenoviral vectors was used, over 80% of target tumor cells could be efficiently transferred. High levels of p53, GM|CSF, and B7|1 expressed in HCC cells transferred with BB|102.Conclusion Adenoviral vectors expressing human wild type p53, GM|CSF and B7|1 genes could be used as an efficient vector to expressing gene |in|interest in HCC cell lines. Our study provides a experimental evidence that the BB |102 might be applicable in clinical gene therapy against HCC.