miR-451a通过MIF调节滋养细胞侵袭能力

miR-451a regulates trophoblast invasion through MIF

目的探讨miR-451a对人绒毛外滋养细胞侵袭能力的调节机制。方法以人工合成的寡核苷酸miR-451a模拟物(miR-451a mimics)转染人绒毛外滋养细胞株HTR-8/SV neo细胞,应用逆转录-实时PCR法检测转染后细胞中miR-451a的表达和巨噬细胞迁移抑制因子(MIF)mRNA表达的变化;应用Western blot法检测转染后细胞中MIF蛋白表达的变化;应用Transwell小室侵袭实验观察转染前后细胞侵袭能力的变化;分别向HTR-8/SVneo细胞中加人重组人巨噬细胞迁移抑制因子(AMIF)和MIF特异性抑制剂ISO-1,应用Western blot法检测细胞中E-钙黏蛋白(E-cadherin)和Vimenthi蛋白表达的变化。结果转染miR-451a mimics后HTR-8/SVneo细胞中miR-451a表达明显升高,MIF mRNA及蛋白表达降低,细胞侵袭能力下降。加入不同药物后,与空白组相比,AMIF组的E-cadherin蛋白表达下降,Vimentin蛋白表达上升;ISO-1组的E-cadherin蛋白表达上升,Vimentin蛋白表达下降。结论转染miR-451a可以下调人绒毛外滋养细胞中MIF的表达,并可能通过上皮间质转化途径(EMT)调控人绒毛外滋养细胞的侵袭能力。

Objective To investigate the regulatory mechanism of miR-451a on the invasion of human villous trophoblasts.Methods Human chorionic villus trophoblast cell line HTR-8/SVneo was transfected with synthetic oligonucleotide miR-451a mimics.Reverse transcription-real-time PCR was used to detect the expression of miR-451a and macrophage migration inhibitory factor(MIF)mRNA.The expression of MIF protein in transfected cells was detected by Western blot.Transwell cell invasion assay was used to observe the changes of cell invasiveness before and after transfection.The recombinant human macrophage migration inhibitory factor(rhMIF)and ISO-1,the specific inhibitors of MIF,were added to HTR-8/SVneo cells.The expression of E-cadherin protein and Vimentin protein in cells was detected by Western blot.Results The level of miR-451a in HTR-8/SVneo cells transfected with miR-451a mimics was significantly increased,the expression of MIF mRNA and protein was decreased,and the cell invasive ability was decreased.Compared with the blank group,E-cadherin protein levels were decreased and Vimentin protein levels increased in the rhMIF group,while increased E-cadherin protein expression and decreased Vimentin protein expression were observed in the ISO-1 group.Conclusions Transfection of miR-451a down-regulates the expression of MIF in human extravillous trophoblasts and may regulate the invasion ability of human villous trophoblast cells through the epithelial-mesenchymal transition pathway.