FasL逆转录病毒转移体系的建立及其在肝癌细胞中的表达

目的　研究Fas/FasL系统的生物学功能，探讨转FasL基因治疗肿瘤的可行性。方法将大鼠FasL全长cDNA亚克隆到逆转录病毒载体pLXSN中，获得FasL正向单拷贝插入子pLXSN/FasL

Development of retroviral vectors containing rat Fas ligand gene and FasL expression mediated by the vectors in hepatocellular carcinoma cells

Objectives　To study the biological functi on of Fas and Fas ligand system and the feasibility of treating tumor with trans fecting FasL gene. Methods　The rat Fas ligand complementa ry DNA was subcloned to the retroviral vector pLXSN to obtain pLXSN/FasL+ reco mbinant with direct inserting then packaged with PA317 amphotropic packaging cel ls, anti-G418 clones were acquired and named as PA317/pLXSN-FasL+ cells. Results　There were FasL gene integration in PA317/ pLXSN -FasL + cells detected by polymerase chain reaction. The titer of virus was 4.7×10 7/L. When the supernatant of the PA317/pLXSNFasL+ cells was used to transfec t hepatocellular carcinoma cells HepG-2, SMMC-7721, CBRH-7919 and RH-35, the F asL positive rate on the hepatocellular carcinoma cells was 97.3%, 99.1%, 99.4% and 72.3% respectively by FCM. The apoptosis in HepG-2 and CBRH-7919 cells with hi gh levels of Fas expression was found. Conclusion　Introduct ion of FasL gene to retroviral vectors was an effective way, which can be used t o induce apoptosis in the cells with high levels of Fas expression and provide t heoretical basis for the gene therapy of tumor.