microRNA-186-5p靶向KLOTHO调控绒毛外滋养细胞侵袭功能的机制研究

Regulation of microRNA-186-5p on invasion of extravillous trophoblast by targeting KLOTHO

目的探讨miR-186-5p通过靶向KLOTHO参与滋养细胞侵袭调控的机制。方法收集2018年6月-2019年6月在武汉大学人民医院妇产科剖宫产的35名子痫前期孕妇及20名正常孕妇的胎盘组织及外周血样本。利用qRT-PCR检测子痫前期及正常胎盘组织miR-186-5p表达水平。取绒毛外滋养细胞系HTR-8/SVneo分组转染,Transwell小室法检测侵袭能力,双荧光素酶报告基因实验验证miR-186-5p与KLOTHO的靶向关系,应用受试者工作特征(ROC)曲线分析外周血miR 186-5p预测子痫前期发生的价值。结果子痫前期孕妇胎盘组织miR-186-5p表达水平明显高于正常胎盘组织,过表达的miR-186-5p显著抑制HTR-8/SVneo细胞侵袭能力,双荧光素酶报告基因实验结果显示,miR-186-5p与KLOTHO直接靶向结合,ROC曲线分析显示,外周血miR-186-5p诊断子痫前期的最佳截断值为2.05,曲线下面积为0.842(95%CI:0.733~0.951),敏感度、特异性分别为80%、76%。结论miR-186-5p可靶向KLOTHO调节滋养细胞侵袭能力,为子痫前期发病机制的研究提供了新线索。

Objective To explore the effect of miR-186-5p on the invasion of extravillous trophoblast by targeting KLOTHO.Methods Placental tissue and peripheral blood samples were collected from 35 preeclampsia pregnant wom⁃en and 20 normal pregnant women who underwent Cesarean section in People’s Hospital of Wuhan University from June 2018 to June 2019.The expression of miR-186-5p in placenta of normal pregnant women and preeclampsia was detected by qRT-PCR.Extravillous trophoblast cell line HTR8/SVneo were manipulated by transfections.The ability of cell invasion was detected by Transwell chamber assay.Dual luciferase reporter assay was used to explore the target⁃ing regulatory relations between miR-186-5p and KLOTHO.The ROC curve was used to analyze the value of serummiR-186-5p in predicting the occurrence of preeclampsia.Results The expression of miR-186-5p in placenta of nor⁃mal pregnant women was significantly lower than that in preeclampsia.Overexpression of miR-186-5p apparently sup⁃pressed invasive ability of HTR8/SVneo cells.The dual luciferase reporter experiment showed that miR-186-5p tar⁃geted KLOTHO.ROC curve analysis showed that the best cut-off value of serum miR-186-5p for the diagnosis of PE was 2.05,the area under the curve was 0.842(95%CI:0.733-0.951),and the sensitivity and specificity were 80%and 76%respectively.Conclusion miR-186-5p targeted regulation of KLOTHO and affected the invasion of extravillous trophoblast in pathogenesis of preeclampsia.