抑制miR-206对子痫前期滋养细胞增殖凋亡及侵袭能力的影响  

Effect of Inhibition of miR-206 on Trophoblast Proliferation,Apoptosis and Invasion in Preeclampsia

作　　者：马会峰[1] 范丽丽[1] 熊丽丽[1] Ma Huifeng;Fan Lili;Xiong Lili(Department of Obstetrics and Gynecology,Kaiyuan Branch,The First Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,China)

机构地区：[1]河南科技大学第一附属医院新区医院妇产科,洛阳471000

出　　处：《成都医学院学报》2024年第4期578-582,共5页Journal of Chengdu Medical College

基　　金：河南省卫生健康委员会医学科技攻关计划联合共建项目(No:LHGJ20200591)。

摘　　要：目的探究抑制miR-206对子痫前期滋养细胞HTR-8/Svneo增殖、凋亡及侵袭能力的影响。方法体外培养人绒毛膜滋养细胞HTR-8/Svneo细胞系,将其分为对照组、缺氧组、缺氧+阴性对照(NC)组和缺氧+miR-206抑制剂(miR-206 inhibitor)组4组。miR-206相对表达量、增殖、凋亡和侵袭情况分别采用实时荧光定量逆转录聚合酶链反应(RT-qPCR)、5-乙炔基-2’-脱氧尿苷(EdU)、Hoechst 33258染色法和Transwell小室测定。E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、细胞周期蛋白D1(Cyclin D1)和半胱氨酸蛋白酶-3(Caspase-3)相对表达量使用蛋白质印迹技术法测定。结果经过转染及氯化钴(CoCl 2)处理后,缺氧组miR-206、E-cadherin和Caspase-3表达及凋亡细胞数均高于对照组,而细胞增殖率、相对侵袭率及Vimentin、N-cadherin和Cyclin D1表达水平均低于对照组(P<0.05)。与缺氧+NC组相比,缺氧+miR-206 inhibitor组miR-206、E-cadherin和Caspase-3表达水平及凋亡细胞数明显降低(P<0.05),而细胞增殖率、相对侵袭率及Vimentin、N-cadherin和Cyclin D1表达水平明显增加(P<0.05)。结论抑制miR-206可能通过促进上皮间质转化(EMT),下调Caspase-3表达水平和上调Cyclin D1蛋白水平来促进缺氧条件下HTR-8/Svneo细胞的增殖和侵袭能力,抑制其凋亡能力。Objective To explore the effects of inhibition of miR-206 on the proliferation,apoptosis and invasion of trophoblast HTR-8/Svneo in preeclampsia.Methods Human chorionic trophoblast HTR-8/Svneo cell line cultured in vitro was divided into 4 groups:control group,hypoxia group,hypoxia+negative control(NC)group and hypoxia+miR-206 inhibitor group.The miR-206 relative expression,proliferation,apoptosis and invasion were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR),5-acetylidene-2'-deoxyuridine(EdU),Hoechst 33258 staining and Transwell assay,respectively.The relative expression levels of E-cadherin,N-cadherin,Vimentin,Cyclin D1 and Cysteine proteinase-3(Caspase-3)were determined by western blotting analysis.Results After the transfection and cobalt chloride(CoCl 2)treatment,the levels of miR-206,E-cadherin and Caspase-3 and the number of apoptotic cells in the hypoxia group were higher than those in the control group,while the cell proliferation rate,relative invasion rate,and the expression levels of Vimentin,N-cadherin and Cyclin D1 were lower than those in the control group(P<0.05).Compared with the hypoxia+NC group,the expression levels of miR-206,E-cadherin and Caspase-3 and the number of apoptotic cells were significantly decreased in the hypoxia+miR-206 inhibitor group,while the cell proliferation rate,relative invasion rate and the expression levels of Vimentin,N-cadherin and Cyclin D1 were significantly increased(P<0.05).Conclusion Inhibition of miR-206 may promote HTR-8/Svneo cell proliferation and invasion and inhibit cell apoptosis under hypoxia condition by promoting epithelial-mesenchymal transition(EMT),down-regulating Caspase-3 and up-regulating Cyclin D1 protein expression.

关 键 词：子痫前期 滋养细胞 miR-206 增殖 凋亡 侵袭 

分 类 号：R714.244[医药卫生—妇产科学]