ELABELA通过调控AKT信号通路影响滋养细胞侵袭的研究

目的 探讨ELABELA(ELA)对妊娠滋养细胞(Bewo细胞)侵袭行为的影响及其可能的作用机制。方法 用siRNA-ELA转染处于对数生长期的Bewo细胞,分为siRNA-ELA转染组,阴性对照组(细胞转染无意义序列)及空白组,细胞转染后于倒置显微镜下观察转染效率,采用实时荧光定量聚合酶链反应(qRT-PCR)检测转染后各组细胞中ELA mRNA表达并明确沉默效率。划痕实验测定各组细胞迁移能力,Transwell实验测定各组细胞的迁移、侵袭能力。蛋白质免疫印迹法检测各组细胞p-AKT、MMP9蛋白表达水平。结果 si-ELA组的ELA表达量明显低于si-NC组及空白组(P<0.001),且si-NC组与空白组差异无统计学意义(P>0.05);siRNA-ELA组细胞的迁移及侵袭能力均明显低于空白组(P<0.01;P<0.01);siRNA-ELA组p-AKT蛋白及MMP9蛋白表达水平明显低于空白组(p-AKT P<0.01;MMP9 P<0.001)。结论 敲低ELA可抑制滋养细胞的侵袭及迁移能力,其机制可能与AKT信号通路有关。

Research on ELABELA affecting trophoblast invasion by regulating AKT signaling pathway

Objective To investigate the effect of ELABELA(ELA) on the invasive behavior of gestational trophoblast cells(Bewo cells) and its possible mechanism.Methods Bewo cells in logarithmic growth phase were transfected with siRNA-ELA,and they were divided into siRNA-ELA transfection group, negative control group(the cells transfected with nonsense sequences) and blank group.The cells were observed transfection efficiency under an inverted microscope after transfection.Real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect the expression of ELA mRNA in each group of cells after transfection, and the silencing efficiency was determined.Scratch test was used to measure the migration ability of cells in each group, and Transwell test was used to measure the migration and invasion ability of cells in each group.Western blot was used to detect the expression levels of p-AKT and MMP9 proteins in each group of cells.Results The expression level of ELA in the si-ELA group was significantly lower than that in the si-NC group and the blank group(P<0.001),and there was no significant difference between the si-NC group and the blank group(P>0.05).The migration and invasion abilities were significantly lower than those in the blank group(P<0.01;P<0.01);the expression levels of p-AKT protein and MMP9 protein in the siRNA-ELA group were significantly lower than those in the blank group(p-AKT P<0.01;MMP9 P<0.001).Conclusion Knockdown of ELA can inhibit the invasion and migration of trophoblast cells, and the mechanism may be related to the AKT signaling pathway.