H19及MMP-2、MMP-9在复发性自然流产患者中的表达及相关性分析

Expression and correlation analysis of H19, MMP-2, MMP-9 in patients with recurrent spontaneous abortion

目的探讨H19及基质金属蛋白酶(MMP)-2、MMP-9在复发性自然流产患者中的表达及相关性。方法体外培养人绒毛外滋养细胞系HTR-8,采用慢病毒感染HTR-8细胞系分别过表达H19及干扰H19的表达,ELISA法检测各组细胞培养上清液中MMP-2及MMP-9蛋白浓度。采用实时荧光定量PCR(qRT-PCR)法检测HTR-8细胞中及复发性自然流产患者绒毛组织中H19、MMP-2、MMP-9 mRNA的表达水平,采用Spearman相关分析H19与MMP-2、MMP-9表达水平的相关性。结果过表达H19后,Lv-pH19组的MMP-2、MMP-9 mRNA表达水平及蛋白浓度显著高于Lv-vector组(P<0.05);干扰H19表达后,Lv-shH19组MMP-2、MMP-9 mRNA表达水平及蛋白浓度显著低于Lv-shcon组(P<0.05)。复发性自然流产患者自然流产次数显著多于对照组(P<0.05)。qRT-PCR结果显示复发性自然流产患者绒毛组织中H19、MMP-2、MMP-9 mRNA的表达水平显著低于对照组(P<0.05)。且H19与MMP-2、MMP-9的表达水平存在正相关(P<0.05)。结论H19可调控滋养细胞MMP-2、MMP-9的表达,早孕期绒毛组织中H19及MMP-2、MMP-9表达异常可能参与自然流产的发生。

Objective To investigate the expression and correlation of H19,matrix metalloproteinase(MMP)-2 and MMP-9 in patients with recurrent spontaneous abortion(RSA).Methods Human extravillous trophoblast cell line HTR-8 was cultured in vitro.Lentivirus was used to infect the HTR-8 cell line to over-express or knockdown the expression of H19.The concentrations of MMP-2 and MMP-9 protein in cell culture supernatant were detected by enzyme linked immunosorbent assay(ELISA).The expression levels of H19,MMP-2 and MMP-9 mRNA in villi of patients with RSA were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Spearman correlation analysis was used to understand the correlation between H19 and the expression levels of MMP-2 and MMP-9.Results After overexpression of H19,the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-ph19 group were significantly higher than those in Lv-vector group(P<0.05);After interfering with the expression of H19,the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-shH19 group were significantly lower than those in Lv-shcon control group(P<0.05).The number of spontaneous abortions in patients with recurrent spontaneous abortion was significantly higher than that in the control group(P<0.05).qRT-PCR showed that the expression levels of H19,MMP-2 and MMP-9 mRNA in villi of patients with RSA were significantly lower than those in the control group(P<0.05).There was a positive correlation between H19 and the expression levels of MMP-2 and MMP-9(P<0.05).Conclusions H19 regulates the expression of MMP-2 and MMP-9 of trophoblast during early pregnancy,and the abnormal expression of H19,MMP-2,and MMP-9 in human first-trimester villous tissues was related with the incidence of early miscarriage.