17β-雌二醇通过microRNA-29b调节NK细胞干扰素γ的分泌  

17β-Estradiol modulates the secretion of NK interferron gamma via microRNA-29b expression

作　　者：李鹏飞[1] 郭葳[1] 赵俊丽[1] 王亚平[1] 胡娅莉[2] 侯亚义[1] 

机构地区：[1]南京大学医学院免疫与生殖生物学实验室,南京210093 [2]南京大学医学院附属鼓楼医院妇产科,南京210008

出　　处：《中国免疫学杂志》2012年第5期418-423,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81072410);江苏省卫生厅科技项目(H200754);江苏省卫生厅"兴卫工程"基金(XK200709);江苏省第五批"六大人才高峰资助"项目资助

摘　　要：目的:研究17β-雌二醇(E2)对人自然杀伤细胞株NK92中干扰素γ(IFN-γ)分泌的影响及其可能的机制。方法:用E2和poly I:C处理NK92细胞,分别在4、8、12和24小时后用实时定量PCR、酶联免疫吸附法和流式细胞术测定NK92细胞中IFN-γ的表达情况,用实时定量PCR检测microRNA-29b(miR-29b)的变化。进一步,构建含有IFN-γ3'UTR的pEGFP-C1质粒,并转染pre-29b和pEGFP-IFN-γ3'UTR质粒至HEK293A细胞中,用流式细胞术检测绿色荧光蛋白(GFP)的变化。结果:E2抑制NK92细胞的IFN-γ分泌和上调miR-29b的表达;miR-29b能够直接靶向调节IFN-γ3'UTR mRNA。结论:E2可能通过调节NK92的miR-29b而影响IFN-γ的分泌水平,进而影响其免疫功能。Objective：To investigate the possible mechanism on the effects of 17β-Estradiol（E2） on human natural killer cell line NK92.Methods：The NK92 were treated with E2 and poly I：C for different time（4,8,12 and 24 hours）.The expression of IFN-γ was measured by qRT-PCR,ELISA and FCM.The expression of miR-29b was determined by qRT-PCR.Meanwhile,the recombinant plasmid pEGFP-IFN-γ 3′ UTR,the pre-29b and pEGFP-IFN-γ 3′ UTR plasmid were constructed and transfected into HEK293A cell by lipofectamine 2000.The expression of GFP was detected by FCM.Results： E2 could down-regulate the secretion of IFN-γ and up-regulate the expression of miR-29b by NK92.Furthermore,miR-29b could modulate the expression of IFN-γ 3′ UTR mRNA directly.Conclusion：The effect of E2 on the immune function of NK cell may be attributed to the secretion of IFN-γ via modulating miR-29b expression.

关 键 词：17Β-雌二醇 NK92 miR-29b IFN-Γ 

分 类 号：R392[医药卫生—免疫学]