MMP-9调控绒毛外滋养细胞sFasL表达的研究

Investigation on expression of sFasL in extravillous trophoblasts regulated by MMP-9

目的:探讨基质金属蛋白酶-9(MMP-9)在绒毛外滋养细胞对可溶性Fas配体(sFasL)表达的调控。方法:化学合成针对MMP-9的siRNA,通过脂质体转染绒毛外滋养细胞株(TEV-1),运用荧光定量逆转录-聚合酶链反应(RealTimeRT-PCR)和ELISA法检测MMP-9siRNA转染前后细胞中MMP-9、FasLmRNA及其蛋白分泌表达的变化。结果:转染后24h,MMP-9siRNA组的MMP-9mRNA表达较对照组明显降低(P<0.05),而FasLmRNA表达较对照组无明显变化(P>0.05)。转染后48h,MMP-9siRNA组的MMP-9蛋白及FasL蛋白的分泌表达较对照组均显著降低(P<0.05)。结论:MMP-9siRNA能特异高效沉默TEV-1的MMP-9mRNA表达,MMP-9蛋白分泌减少的同时也降低了sFasL蛋白的分泌。绒毛外滋养细胞表达的MMP-9可调节sFasL的表达,从而影响内皮细胞凋亡,可能参与早期胎盘血管的重建过程。

Objective：To explore the expression of soluble Fas ligand（sFasL）regulated by matrix metalloproteinase-9（MMP-9）in extravillous trophoblasts.Methods：siRNA targeted to MMP-9 were chemosynthesized,then transfected into extravillous trophoblasts cell line（TEV-1）by lipofectamine,the changes of expression levels of MMP-9,FasL mRNA and proteins before and after transfection were detected by real-time quantitative reverse transcription-polymerase chain reaction（real time RT-PCR）and ELISA.Results：24 hours later after transfection,the expression level of MMP-9 mRNA in MMP-9 siRNA group was significantly lower than that in control group（P0.05）,but there was no significant difference in expression level of FasL mRNA between the two groups（P0.05）.48 hours later after transfection,the expression levels of MMP-9 protein and FasL protein in MMP-9 siRNA group were significantly lower than those in control group（P0.05）.Conclusion：MMP-9 siRNA can silence the expression of MMP-9 mRNA specially and efficiently,decrease the expression levels of MMP-9 protein and FasL protein.MMP-9 expressed by extravillous trophoblasts can regulate the expression of sFasL,affect the apoptosis of endothelial cells and may participate in the reconstruction of early placental vessels.