miR-18a对人正常滋养细胞中MMP-2、MMP-9表达的影响

Effect of MiR-18a on the Expression of MMP-2 and MMP-9 in Human Normal Trophoblast Cells

目的:研究子痫前期(pre-eclapmpsia,PE)相关微小RNA-18a(miR-18a)对人正常滋养细胞(HTR-8细胞)中基质金属蛋白酶2(MMP-2)、MMP-9的影响。方法:按照HTR-8细胞中miR-18a的表达情况分为3组(每组重复5次):miR-18a过表达组、miR-18a抑制组、阴性对照组;实时定量聚合酶链反应(RT-qPCR)检测转染后HTR-8细胞中miR-18amRNA的表达量;RT-qPCR和蛋白质印迹(Western blotting)检测转染后HTR-8细胞中MMP-2、MMP-9mRNA和蛋白表达量。结果:miR-18a过表达组miR-18amRNA表达量比阴性对照组升高,miR-18a抑制组miR-18amRNA表达量比阴性对照组降低,差异均有统计学意义(均P<0.001)。miR-18a抑制组MMP-2、MMP-9mRNA和蛋白表达量比阴性对照组降低,差异均有统计学意义(均P<0.001)。结论:miR-18a影响人正常滋养细胞中MMP-2、MMP-9在mRNA及蛋白水平的表达,可能通过该途径参与对滋养细胞浸润能力的调控。

Objective:To investigate the effect of pre-eclapmsia related miR-18a on the expression of matrix metallopeptidase 2(MMP-2)and MMP-9 in human normal trophoblast cells(HTR-8).Methods:According to the expression of miR-18a in HTR-8 cells,it was divided into 3 groups(5 times in each group):miR-18a overexpression group,miR-18a inhibition group,negative control group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-18a mRNA in HTR-8 cells after transfection.The mRNA and protein levels of MMP-2 and MMP-9 in HTR-8 cells were detected by RT-qPCR and Western blotting after transfection.Results:The expression of miR-18a mRNA in miR-18a overexpression group was higher than that in negative control group(P<0.001).The expression of miR-18a mRNA in miR-18a inhibition group was lower than that in negative control group(P<0.001).The expression of MMP-2 and MMP-9 mRNA and protein in the miR-18a inhibition group was lower than that in the negative control group,and the difference was statistically significant(both P<0.001).Conclusions:MiR-18a could affect the expression of MMP-2 and MMP-9 in human normal trophoblast cells at mRNA and protein levels,and may be involved in the regulation of trophoblast infiltration through this pathway.