中国肿瘤生物治疗杂志2018，Vol.25Issue(1)：73-78,6.DOI:10.3872/j.issn.1007-385x.2018.01.013
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
摘要
Abstract
Objective:To investigate the associations of MHC class Ⅰ chain-related molecule A (MICA) gene polymorphism with susceptibility 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 ofperforin(PFN) and granzymes B(Gzm B) were measured by ELISPOT assay.Results:DNA sequencing result showed that MICA*008/A5.1 and MICA*001/A4 were expressed in MCF-7 cells,MICA*019/A5 and MICA*002/A9 were expressed in both MDA-MB-231 and SK-BR-3 cells while MICA*010/A5 was expressed in MDA-MB-435S cells.After the MICA recombinant 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 cytotoxicity 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 difference 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Key words
breast cancer cell/MHC class I chain-related molecule A (MICA) gene/polymorphism/natural killer cell (NK cell)分类
医药卫生引用本文复制引用
陈淑萍,周智锋,林万松,李洁羽,刘枋,叶韵斌..MICA基因多态性与乳腺癌细胞对NK细胞杀伤的敏感性[J].中国肿瘤生物治疗杂志,2018,25(1):73-78,6.基金项目
国家临床重点专科建设项目资助(国卫办医函[2013]544号) （国卫办医函[2013]544号）
福建省自然科学基金资助项目(No.2015J01433). Project supported by the National Clinical Key Specialty Construction Program of China (Letter of the National Health Office [2013] 544),and the Natural Science Foundation of Fujian Province(No.2015J01433) （No.2015J01433）
