MICA基因多态性与乳腺癌细胞对NK细胞杀伤的敏感性  

Associations of MICA gene polymorphism with susceptibility of breast cancer cells to NK cell-mediated cytotoxicity

作　　者：陈淑萍[1] 周智锋[1] 林万松[1] 李洁羽[1] 刘枋[1] 叶韵斌[1] 

机构地区：[1]福建省肿瘤医院,福建医科大学附属肿瘤医院肿瘤免疫学研究室,福建省肿瘤转化医学重点实验室,福建福州350014

出　　处：《中国肿瘤生物治疗杂志》2018年第1期73-78,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家临床重点专科建设项目资助(国卫办医函[2013]544号)：福建省自然科学基金资助项目(No.2015J01433).

摘　　要：目的:探讨MHC-Ⅰ类链相关分子A(MHC class I chain-related molecule A,MICA)多态性与乳腺癌细胞对NK细胞杀伤敏感性的关系。方法:测序分析乳腺癌细胞系MCF-7、MDA-MB-231、MDA-MB-435S和SK-BR-3的MICA等位基因,用Western blotting和流式细胞术检测MICA重组表达载体转染293T细胞(分别命名为p MCFA5.1、p MCFA4、p231A5R、p231A9和p435A5P)MICA蛋白的表达水平,用LDH法检测NK细胞对转染MICA的293T细胞的杀伤活性,酶联免疫斑点法检测NK细胞穿孔素、颗粒酶B分泌水平。结果:MCF-7细胞表达MICA*008/A5.1和MICA*001/A4,MDA-MB-231和SK-BR-3细胞均表达MICA*019/A5和MICA*002/A9,MDA-MB-435S细胞表达MICA*010/A5。转染MICA后,p MCFA5.1组293T细胞MICA蛋白的表达水平最低(P<0.05),p435A5P组次之(P<0.05),p MCFA4组、p231A5R组和p231A9组表达水平较高(均P<0.05)。NK细胞对转染MICA的293T细胞杀伤活性及穿孔素、颗粒酶B分泌:p435A5P组对NK细胞杀伤的敏感性最低(P<0.05),穿孔素、颗粒酶B分泌水平最低(均P<0.05);p MCFA5.1、p MCFA4、p231A5R和p231A9各组之间比较差异无统计学意义(P>0.05)。结论:MICA基因多态性与乳腺癌细胞对NK细胞杀伤的敏感性密切相关。Objective： To investigate the associations of MHC class I chain-related molecule A （MICA） gene polymorphism with sus- ceptibility of breast cancer cells to NK cell-mediated cytotoxicity. Methods： MICA alleles of breast cancer cell lines （MCF-7, MDA- MB-231, MDA-MB-435S and SK-BR-3） were analyzed by DNA sequencing. MICA protein expression in 293T cells transfected with MICA recombinant expression vectors （namely pMCFA5.1, pMCFA4, p231A5R, p231A9 and p435A5P） was detected by Western blotting and Flow cytometry; the cytotoxicity of NK cells against the above 293T cells were measured by lactate dehydrogenase （LDH） assay and the release of perforin（PFN） and granzymes B （Gzm B） were measured by ELI SPOT assay. Results： DNA sequenc- ing result showed that MICA＊OO8/A5.1 and MICA＊OO1/A4 were expressed in MCF-7 cells, MICA＊O19/A5 and MICA＊OO2/A9 were ex- pressed in both MDA-MB-231 and SK-BR-3 cells while MICA＊OIO/A5 was expressed in MDA-MB-435S cells. After the MICA recom- binant expression vectors were transfected into 293T cells, the level of MICA were the lowest in pMCFA5.1 group （P〈0.05）, following after p435A5P group （P〈0.05）, and highly expressed in the pMCFA4, p231AR and p231A9 groups （P〈 0.05）. NK cell-mediated cyto- toxicity and the release of PFN and Gzm B： NK cell-mediated cytotoxicity were the lowest in p435A5P group （P〈0.05）, and the ability of inducing NK cells to release PFN and Gzm B was also the lowest in p435A5P group （P〈0.05）, but there were no statistical differ- ence among the other transfected groups （P〉0.05）. Conclusion： MICA gene polymorphism is closely associated with susceptibility of breast cancer cells to NK cell-mediated cytotoxicity.

关 键 词：乳腺癌细胞 MICA基因 多态性 自然杀伤细胞 

分 类 号：R737.9[医药卫生—肿瘤] R730.54[医药卫生—临床医学]