MAGE-3基因稳定转染的HHCC细胞系的建立及其mRNA的表达  

Establishment of an HHCC cell line stably transfected by tumor antigen MAGE-3 gene and its mRNA expression

作　　者：张秀敏[1] 隋延仿[1] 司少艳[1] 李侠[1] 黄杨[1] 胡沛臻[1] 葛伟[1] 

机构地区：[1]第四军医大学基础部病理学教研室,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2006年第4期450-451,453,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30271464);全军医药卫生科研基金重点项目(01Z084)

摘　　要：目的:构建肿瘤抗原MAGE-3基因的真核表达载体,建立MAGE-3基因稳定转染的人肝细胞癌细胞系(humanhepatocellularcarcinomacellline,HHCC)。方法:利用分子生物学方法,构建带有MAGE-3基因和增强型绿色荧光蛋白(EGFP)报告基因的重组真核表达质粒pIRES2-EGFP-MAGE-3,经脂质体法转染HHCC后,用G418筛选阳性克隆。在荧光显微镜下观察HHCC中EGFP的表达,用RT-PCR法检测HHCC中MAGE-3mRNA的表达。结果:成功地构建了重组真核表达载体pIRES2-EGFP-MAGE-3。以其转染HHCC后,经荧光显微镜及RT-PCR分别检测到MAGE-3mRNA转录和EGFP的表达。结论:建立了1株可稳定转染MAGE-3基因的肝细胞癌HHCC,为进一步应用该基因进行肝癌的免疫治疗提供了实验依据。AIM： To construct the eukaryotic expression vector of tumor antigen MAGE-3 and establish human hepatocellular carcinoma cell line （HHCC） expressing MAGE-3. METHODS： The MAGE-3 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-3 plasmid. The recombinant plasmid pIRES2-EGFP-MAGE-3 was transfected into HHCC cells by lipofectamine, and then the positive clones were screened by G418. The expression of enhanced green fluorescent protein （EGFP） and MAGE-3 mRNA in positive clones were detected by fluorescence microscope and RT-PCR, respectively. RESULTS： The eukaryotic expression vector pIRES2-EGFP-MAGE-3 was successfully constructed. The expression of EGFP was found by fluorescence microscope detection and MAGE-3 mRNA transcription was detected by RT-PCR in the positive clones. CONCLUSION： The stable MAGE-3-transfected HHCC cell line is successfully established, which will provide experimental basis for further study on immunotherapy for hepatocellular carcinoma using MAGE-3 as target antigen.

关 键 词：黑色素瘤抗原-3 基因转染 绿色荧光蛋白 肿瘤 免疫治疗 

分 类 号：R329.11[医药卫生—人体解剖和组织胚胎学]