miR-152 通过HLA-G / KIR2DL4 通路对人正常滋养细胞中基质金属蛋白酶2、9 表达的影响  

The effect of miR-152 on the expression of matrix metalloproteinase 2and 9 in human normal trophoblast cells through the HLA-G /KIR2DL4 pathway

作　　者：杨洋 马媛[3] 宋姗姗 陈宥艺[1] 张建华 Yang Yang;Ma Yuan;Song Shanshan;Chen Youyi;Zhang Jianhua(Reproductive Medicine Center,Xi'an People's Hospital(Xi'an Fourth Hospital),Xi'an Shaanxi 710004;Xi'an Medical University,Xi'an Shaanxi 710021;Department of Obstetrics and Gynecology,Tangdu Hospital,Chinese People's Liberation Army Air Force Military Medical University,Xi'an Shaanxi 710038;2nd Department of Obstetrics,The Second Affiliated Hospital of Xi'an Medical University,Xi'an Shaanxi 710038,P.R.China)

机构地区：[1]西安市人民医院(西安市第四医院)生殖医学中心,陕西西安710004 [2]西安医学院,陕西西安710021 [3]中国人民解放军空军军医大学唐都医院妇产科,陕西西安710038 [4]西安医学院第二附属医院产二科,陕西西安710038

出　　处：《中国计划生育和妇产科》2022年第7期84-88,I0001,共6页Chinese Journal of Family Planning & Gynecotokology

基　　金：国家自然科学基金项目(项目编号:81901510);西安市科技计划项目(项目编号:20YXYJ0005(6));陕西省自然科学基础研究计划(项目编号:2020JM-615)。

摘　　要：目的研究miR-152通过其靶基因人类白细胞抗原G(human leukocyte antigen-G,HLA-G)与蜕膜NK细胞(decidual natural killer,dNK)表面杀伤细胞免疫球蛋白样受体(killer Ig-like receptors,KIR)2DL4之间通路,对人正常滋养细胞(HTR-8)中基质金属蛋白酶(matrix metalloproteinases,MMP)2、9表达的影响。方法收集2020年9月~10月西安市第四医院妇产科门诊人工流产患者正常早孕蜕膜组织(n=5),分选、纯化dNK;构建其与过表达和抑制miR-152后的HTR-8细胞共培养体系,在共培养中封闭dNK表面杀伤细胞免疫球蛋白样受体(killer Ig-like receptors,KIR)KIR2DL4。实时定量PCR(Real-time quantitative PCR,qRT-PCR)检测miR-152转染效率;qRT-PCR、蛋白免疫印迹检测转染miR-152及干预HLA-G/KIR2DL4通路后HTR-8中MMP2、9的表达,Transwell实验检测HTR-8浸润能力的变化情况。结果miR-152在HTR-8细胞中过表达和抑制效果显著;在抑制miR-152表达的共培养组HTR-8中MMP2、MMP9在mRNA及蛋白水平表达升高,且细胞浸润能力增强;过表达miR-152且封闭KIR2DL4共培养组HTR-8浸润能力显著降低,MMP2、MMP9在mRNA及蛋白水平表达降低,差异均有统计学意义(P<0.05)。结论miR-152能够通过HLA-G/KIR2DL4通路影响HTR-8细胞中MMP2、MMP 9表达。Objective To study the pathway of miR-152 through its target gene human leukocyte antigen G and the killer cell immunoglobulin-like receptor(KIR) KIR2 DL4 on the surface of decidual NK cells, which could affect the expression of matrix metalloproteinase(MMP) 2 and 9 in human normal trophoblast cells.Methods Collected human normal early pregnancy decidua tissue of abortion patients in Obstetrics and Gynecology Cliaic of Xi’an Forth Hospital from September 2020 to October 2020(n=5),sort and purify dNK;construct a co-cultivation system with HTR-8 cells after surface and miR-152 inhibition, and block dNK surface killer cell immunoglobulin in the co-culture.Real-time quantitative PCR(qRT-PCR) detects the transfection efficiency of miR-152;qRT-PCR and Western-blot were used to detect the expression of MMP2 and 9 in HTR-8 after tranfection of miR-152 and intervention HLA-a/KIR2 DL4 pathway.Trans-well experiment was used to detect the changes of HTR-8 infiltration capacity.Results Overexpression and inhibitory effect of miR-152 was significant in HTR-8 cells;In the co-culture group HTR-8 that inhibited miR-152 expression, the expression of MMP2 and 9 increased at mRNA and protein levels, and the cell infiltration ability was enhanced;the HTR-8 infiltration ability of the co-culture group that overexpressed miR-152 and blocked KIR2 DL4 was significantly reduced, and expression of MMP2 and 9 were reduced at mRNA and protein levels.Conclusion miR-152 can affect the expression of MMP2 and MMP 9 in HTR-8 cells through the HLA-G/KIR2 DL4 pathway.

关 键 词：微小RNA 蜕膜NK细胞 滋养细胞 浸润能力 基质金属蛋白酶 

分 类 号：R714.24[医药卫生—妇产科学]