子痫前期相关miR-210通过下调钾离子通道调控因子1(KCMF1)抑制滋养细胞的侵袭

Preeclampsia-related miR-210 inhibits invasion of trophoblast cells by down-regulating potassium channel regulator 1(KCMF1)

目的研究子痫前期(PE)相关miR-210通过调控钾离子通道调控因子1(KCMF1)影响滋养细胞侵袭功能的机制。方法选取24例PE患者及25例同期健康产妇,实时荧光定量PCR检测其血清、胎盘组织miR-210相对表达量。采用脂质体转染法将miR-210抑制物(miR-210 inhibitor)及阴性对照、miR-210模拟物(miR-210 mimimc)及阴性对照转染至HTR8/Svneo细胞并设置空白对照组。转染48 h后,采用荧光显微镜观察转染效率;Transwell^(TM)小室实验测定各组细胞侵袭能力;实时荧光定量PCR检测HTR8/Svneo细胞miR-210、KCMF1 mRNA水平、Western blot法检测KCMF1蛋白相对表达量;双荧光素酶实验检验miR-210与KCMF1的靶向关系。结果PE组血清及胎盘组织miR-210相对表达量均高于对照组。转染后48 h,各组细胞转染效率均>80%;与其他组比较,miR-210 mimics组穿膜细胞数减少,miR-210相对表达量增加,KCMF1 mRNA及蛋白相对表达量降低,相对荧光素酶活性降低;miR-210 inhibitor组miR-210相对表达量降低。结论PE血清及胎盘中miR-210异常高表达,上调miR-210引起KCMF1表达下调,抑制滋养细胞侵袭。

Objective To explore the mechanism of preeclampsia(PE)-related miR-210 affecting the invasion of trophoblast cells by regulating potassium channel regulator 1(KCMF1).Methods Twenty-four PE patients and 25 healthy parturients during the same period were selected,and the relative expression of miR-210 in serum and placenta tissue was detected by real-time fluorescent quantitative PCR.The liposome transfection method was used to transfect miR-210 inhibitor and negative control,miR-210 mimic and negative control into HTR8/Svneo cells,meanwhile a blank control group was set.Fluorescence microscope was used to observe the transfection efficiency after 48 hours of transfection.Transwell^(TM) chamber test was used to determine the invasive ability of each group.Real-time fluorescent quantitative PCR was used to detect the relative expression of miR-210 and KCMF1 mRNA in HTR8/Svneo cells.Western blot analysis was used to detect the relative expression of KCMF1 protein.Double luciferase experiment was used to test the targeting relationship between miR-210 and KCMF1.Results The relative expression of miR-210 in serum and placenta tissue of PE group was higher than that of normal control group.At 48 hours after transfection,the transfection efficiency of each group was more than 80%.The number of transmembrane cells in the miR-210 mimic group was less than that of other groups.The relative expression of miR-210 in miR-210 mimic group was higher than that of other groups.The relative expression of KCMF1 mRNA and protein,the relative luciferase activity were lower than those of other groups.The relative expression of miR-210 in the miR-210 inhibitor group was reduced.Conclusion The expression of miR-210 is abnormally high in PE serum and placenta.Up-regulation of miR-210 can cause down-regulation of KCMF1 expression and inhibit trophoblast cell invasion.