云南宣威肺腺癌细胞XWLC-05转染HLA-A0201的抗肿瘤研究  

The Role of HLA-A*0201 in Tumor Treatment by Transfected into Xuanwei Lung Adenocarcinoma Cell Line XWLC-05

作　　者：邵文萍[1] 罗劭蕾 郝萍[1] 马丽菊[1] 唐睿株 闫凤彩[1] 

机构地区：[1]昆明医科大学第一附属医院临床医学实验研究科,云南昆明650032 [2]云南大学医学院,云南昆明650091

出　　处：《昆明医科大学学报》2015年第4期10-14,共5页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(81160276);云南省科技厅-昆明医科大学联合专项基金资助项目(2011FB82)

摘　　要：目的在云南宣威肺腺癌细胞XWLC-05中转染并表达外源性云南肺癌患者白细胞抗原(human leukocyte antigen,HLA)等位基因高频位点HLA-A*0201基因,以诱导肿瘤抗原特异性的细胞毒作用.方法采用序列特异性引物扩增(PCR-SSP)法对XWLC-05细胞行HLA-DNA分型,脂质体转染法将真核质粒pcDNA3.1-HLA-A*0201导入XWLC-05细胞并用G418筛选阳性克隆,淋巴细胞分离液提取健康人外周血单个核细胞(PBMNCs)诱导培养为LAK细胞,免疫磁珠分选法提取出CD8+T淋巴细胞(CTLs),MTT法测试肿瘤特异性的细胞毒效应.结果筛选出稳定表达HLA-A*0201基因的阳性克隆,免疫激活并分选出HLA-A*0201阳性的CTLs,该细胞对转染了HLA-A*0201基因的肿瘤细胞有更强的杀伤活性.结论 HLA-A*0201基因低表达或缺失,在导致肿瘤细胞免疫逃逸方面起重要作用,HLA-A*0201基因可诱导出HLA-A*0201限制的肿瘤特异性T淋巴细胞杀伤效应.Objective HLA-A＊0201 was transfected and expressed in Yunnan Xuanwei lung cancer cell line XWLC-05 in order to induce the tumor antigen-specific cytotoxicity. Methods XWLC-05 cells were HLA-DNA typed using sequence-specific primers PCR （PCR-SSP） , and the cells were transfected with eukaryotic expressing plasmid pcDNA3.1-HLA-A＊0201 or the control vector by liposome transfection. Positive clones were screened with G418. Healthy human peripheral blood mononuclear ceils （PBMNCs） were extracted using lymphocyte separation medium, and them were activated or become LAK cells with PHA or rhIL-2, respectively. The CD8＋ T lymphocytes （CTLs） were extracted from them by immunomagnetic beads, respectively. MTr was used to assay the effects of tumor-specific cytotoxicity. Results The positive clones stably expressing HLA-A＊0201 were gotten, the CTLs expressing HLA-A＊0201 were activated and sorted from PBMNCs of healthy human, and these CTLs showed powerful ability to kill the cancer cells transfected with HLA-A＊0201. Conclusion Deletion or weak expression HLA-A＊0201 likely plays an important role in tumor cells escaping from host immune defenses, and HLA-A＊0201-restricted tumor-specific T lymphocyte killing effect could be induced by HLA-A＊0201.

关 键 词：HLA-A＊0201 基因转染 肿瘤细胞 LAK细胞 

分 类 号：R730.3[医药卫生—肿瘤]