miR-204在子痫前期患者胎盘组织中的表达及对滋养细胞增殖和侵袭功能影响观察

Expression of miR-204 in placenta tissues of preeclampsia patients and its influence on trophoblast proliferation and invasion

目的观察miR-204在子痫前期(PE)患者胎盘组织中的表达变化及其对滋养细胞增殖、侵袭功能的影响。方法采用实时荧光定量PCR(qRT-PCR)检测60例子痫前期患者和30例正常妊娠孕妇胎盘组织中的miR-204。采用生物学信息法预测miR-204的可能作用靶基因。采用脂质体转染法转染miR-204模拟物至人绒毛膜滋养细胞HTR-8/SVneo,CCK8实验检测细胞增殖能力,Transwell小室实验检测细胞侵袭能力,qRT-PCR和Western blotting检测细胞基质金属蛋白酶(MMP)-9 mRNA和蛋白。结果 PE患者和正常孕妇胎盘组织中miR-204的相对表达量分别为2.40±0.16、1.02±0.05,两者相比,P<0.01;重度PE和轻度PE患者胎盘组织中miR-204的相对表达量分别为2.84±0.19、1.96±0.13,两者相比,P<0.01。miR-204作用靶基因可能是MMP-9。与对照细胞相比,转染miR-204模拟物的HTR-8/SVneo细胞的增殖能力和侵袭能力明显降低(P均<0.01)、MMP-9 mRNA和蛋白的表达明显下降(P均<0.01)。结论 miR-204在PE患者胎盘组织中表达明显增高。miR-204能够通过下调MMP-9的表达使滋养细胞增殖和侵袭能力下降,与PE的发病有关。

Objective To observe the changes in miR-204 expression in placenta tissues of preeclampsia(PE)patients,as well as its effect on trophoblast proliferation and invasion.Methods The miR-204 expression in the placenta tissues of 60 PE patients and 30 normal pregnant women was detected by using quantitative real-time fluorescent PCR(qRT-PCR),and the potential target gene of miR-204 was predicted through bioinformatics.The miR-204 mimics was transfected into the human villous trophoblast HTR-8/SVneo through lipofection transfection,the cell proliferation capacity was detected through CCK8 assay,cell invasion capacity was determined through Transwell assay,and the matrix metalloproteinase(MMP)-9 mRNA and protein expression was detected by using qRT-PCR and Western blotting,respectively.Results The relative expression of miR-204 in the placenta tissues of PE patients and normal pregnant women was 2.40±0.16 and 1.02±0.05,respectively,and the difference between two groups was statistically significant(P<0.01).The relative expression of miR-204 in the placenta tissues of severe PE and mild PE patients was 2.84±0.19 and 1.96±0.13,respectively,with statistically significant difference(P<0.01).MMP-9 might be the potential target gene of miR-204.Compared with the control cells,HTR-8/SVneo cells transfected with miR-204 mimics had remarkably reduced proliferation and invasion capacities(all P<0.01),along with down-regulated MMP-9 mRNA and protein expression(all P<0.01).Conclusion The miR-204 expression is notably up-regulated in the placenta tissues of PE patients.The miR-204 can down-regulate MMP-9 expression to lower the proliferation and invasion capacities of trophoblasts,which is related to the genesis of PE.