microRNA125a-3p对滋养层细胞功能的调控作用及机制  

Regulation and mechanism of microRNA125a-3p on trophoblast cell function

作　　者：刘倩[1] 张琦[1] 谢青贞[1] LIU Qian;ZHANG Qi;XIE Qing-zhen(Center for Reproductive Medicine,Renmin Hospital of Wuhan University,Wuhan 430060)

机构地区：[1]武汉大学人民医院生殖医学中心,武汉430060

出　　处：《生殖医学杂志》2023年第2期260-268,共9页Journal of Reproductive Medicine

基　　金：国家自然科学基金青年项目(81801452)。

摘　　要：目的观察microRNA125a-3p(miR-125a-3p)在滋养层细胞中的表达,探讨其对滋养层细胞增殖、侵袭和凋亡的调控及机制。方法荧光实时定量PCR检测人滋养层细胞系HTR-8/SVneo、绒癌细胞系JAR和JEG-3中miR-125a-3p的表达情况。以HTR-8/SVneo和JEG-3细胞为实验对象,分为3组:空白对照组(CK组),未做任何处理;阴性对照组(NC组),转染NC-inhibitor;实验组(inhibitor组),转染miR-125a-3p inhibitor。以Transwell、流式细胞仪、CCK8法分别检测细胞的侵袭、凋亡及增殖能力。Western blot检测Fyn蛋白表达情况及ERK1/2、STAT3磷酸化水平。荧光实时定量PCR检测Fyn mRNA水平,免疫共沉淀法检测Fyn活性水平。结果miR-125a-3p mRNA表达水平在HTR-8/SVneo、JAR和JEG-3细胞中依次降低,两两比较均有统计学差异(P<0.01)。抑制HTR-8/SVneo和JEG-3中miR-125a-3p后,细胞的凋亡水平明显降低,侵袭和增殖能力均明显升高(P<0.05);Fyn mRNA和蛋白的表达及活性水平均明显升高(P<0.05);ERK1/2及STAT3的磷酸化水平均不同程度增加(P<0.05)。结论本研究首次在滋养层细胞中检测到miR-125a-3p的表达。miR-125a-3p通过作用于Fyn和ERK1/2-STAT3信号通路可抑制滋养层细胞的增殖、侵袭,促进其凋亡。Objective:To observe the microRNA125a-3p(miR-125a-3p)expression in trophoblast cell,and to explore its roles in trophoblast cell migration,proliferation and apoptosis,and the related mechanism.Methods:Quantitative real-time PCR was used to detect the expression of miR-125a-3p in HTR-8/SVneo,choriocarcinoma cell line JAR and JEG-3.HTR-8/SVneo and JEG-3 cells were divided into three groups:blank control group(CK group)without any treatment;negative control group(NC group),transfected with NC inhibitor;and the experimental group(inhibitor group)was transfected with miR-125a-3p inhibitor.The invasion,proliferation and apoptosis of trophoblast cell were respectively assessed by using Transwell assay,flow cytometry and cell counting kit-8 assay.Fyn protein expression and ERK1/2,STAT3 phosphorylation levels were detected by Western blot.Fyn mRNA expression were evaluated by quantitative real-time PCR.Fyn activity was evaluated through immunoprecipitation.Results:The level of miR-125a-3p significantly decreased in order from HTR-8/SVneo,JAR to JEG-3(P<0.01).After inhibiting miR-125a-3p in HTR-8/SVneo and JEG-3,the apoptosis level of cells was significantly reduced,and the invasion and proliferation ability were significantly increased(P<0.05).Meanwhile,Fyn expression and activity were significantly increased(P<0.05).Moreover,the phosphorylation levels of ERK1/2 and STAT3 increased in varying degrees(P<0.05).Conclusions:Our study firstly detects the expression of miR-125a-3p in trophoblast cell.miR-125a-3p inhibits the proliferation and invasion of trophoblast cell and promotes its apoptosis by acting on Fyn and ERK1/2-STAT3 signaling pathways.

关 键 词：miR-125a-3p 滋养层细胞 酪氨酸激酶 细胞外信号调节激酶(ERK1/2) 信号传导和转录激活因子3(STAT3) 

分 类 号：R34[医药卫生—基础医学]