中文摘要：

目的 构建表达人类白细胞抗原-E（HLA—E）基因慢病毒载体，探讨慢病毒介导HLA—E基因在肿瘤免疫中的意义。方法 2006年10月至2007年11月在中山大学附属第三医院肝移植中心应用pGC—E1-GFP—HLA-E慢病毒载体转导肝癌HepG2细胞，检测慢病毒载体的转导效率、细胞内HLA—E信使核糖核酸（mRNA）及蛋白质水平的表达。结果 pGC—E1-GFP—HLA—E慢病毒载体转导肝癌HepG2细胞72h的转导效率为（95.0±1．3）％；通过RT—PCR检测24h后细胞内HLA—EmRNA水平是肝癌HepG2细胞的（8．73±1．05）倍，72h后为（293．48±42.01）倍，差异具有统计学意义（P〈0．01）。细胞免疫组化显示转导HLA—E后细胞内的蛋白水平明显增强。结论 慢病毒载体系统能够使转导的基因在靶细胞中得到稳定表达，是基因治疗中的理想载体。

英文摘要：

Objective To construct the lentiviral expression vector of HLA-E gene and investigate its significance for further study on tumor immunity. Methods After the lentiviral expression vector of HLA-E gene （pGC-E1-GFP-HLA-E） was transduced HepG2 cell lines,the transduction efficiency, the HLA-E mRNA level by RT-PCR and the expression of HLA-E by immunohistochemistry were detected between October 2006 and November 2007 in the Liver Transplautation Center of Third Affiliated Hospital of Sun Yat-sen University. Results The transduction efficiency of HepG2 cell lines using the lentiviral vectors was 95.0% ± 1.3% which allowing the stable expression of HLA-E after 72 hr. The HLA-E mRNA level in vitro was detected more 8. 73 ± 1.05 times and 293.48±42. 01 times than normal level by RT-PCR （P 〈 0. 01 ）. The expression of HLA-E by immunohistochemistry was displayed significant enhancement. Conclusion The lentiviral expression vector is ideal vetor of gene therapy and made genes transducted stable expression in target cell.