HLA-E基因慢病毒载体构建及其在肝癌HepG2细胞中的表达

张彤1,方天翎2,李华1,聂常富1,蔡常洁1,许赤1,汪根树1,李玺1,易慧敏1,姜楠1,陈规划1

中国实用外科杂志 ›› 2008, Vol. 28 ›› Issue (03) : 213-215.

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PDF(2175 KB)
中国实用外科杂志 ›› 2008, Vol. 28 ›› Issue (03) : 213-215.
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HLA-E基因慢病毒载体构建及其在肝癌HepG2细胞中的表达

  • 张彤1,方天翎2,李华1,聂常富1,蔡常洁1,许赤1,汪根树1,李玺1,易慧敏1,姜楠1,陈规划1
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Construction of recombinant lentivirol expression vector carrying human HLA-E gene and Its expression in HepG2 cells

  • ZHANG Tong,FANG Tian-ling,LI Hua,et al.
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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E mRNA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。

Abstract

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.

关键词

慢病毒 / 人类白细胞抗原-E / 基因表达

Key words

lentivirus / HLA-E / gene expression

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张彤1,方天翎2,李华1,聂常富1,蔡常洁1,许赤1,汪根树1,李玺1,易慧敏1,姜楠1,陈规划1. HLA-E基因慢病毒载体构建及其在肝癌HepG2细胞中的表达[J]. 中国实用外科杂志. 2008, 28(03): 213-215
ZHANG Tong,FANG Tian-ling,LI Hua,et al.. Construction of recombinant lentivirol expression vector carrying human HLA-E gene and Its expression in HepG2 cells[J]. Chinese Journal of Practical Surgery. 2008, 28(03): 213-215

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