Gadd45α mRNA和蛋白在子痫前期胎盘组织中表达及转染siRNA对滋养细胞侵袭能力及侵袭相关分子的影响

Expression of Gadd45α mRNA and protein in placenta of preeclampsia and effect of transfection of siRNA on invasive ability of trophoblasts and invasion-related molecules

目的分析生长阻滞和DNA损伤45α(Gadd45α)蛋白和mRNA在子痫前期(PE)胎盘组织中的表达情况及转染siRNA对滋养细胞侵袭能力及侵袭相关分子的影响。方法回顾性选取咸宁市中心医院收治的40例PE胎盘组织为病例组,30例正常胎盘组织为对照组。通过免疫印迹法对两组Gadd45α蛋白表达水平进行检测,并用实时荧光定量PCR法对两组Gadd45αmRNA的相对表达量进行检测。对滋养细胞HTR8/Svneo进行培养,以转染阴性对照的siRNA为对照siRNA组,转染Gadd45α的siRNA为Gadd45α-siRNA组。在转染Gadd45α的siRNA后对滋养细胞侵袭数量和侵袭相关分子基质金属蛋白酶(MMPs)的表达水平进行检测。结果相比对照组,病例组Gadd45α蛋白表达水平明显升高(P<0.01),Gadd45αmRNA的相对表达量明显上调(P<0.01)。相比对照siRNA组,Gadd45α-siRNA组滋养细胞HTR8/Svneo中Gadd45α蛋白表达水平显著下降(P<0.01),Gadd45αmRNA的相对表达量亦显著下调(P<0.01)。随着转染siRNA时间的增加,对照siRNA组与Gadd45α-siRNA组细胞侵袭数量明显增多(P<0.05);相比对照siRNA组,Gadd45α-siRNA组同时间点滋养细胞HTR8/Svneo侵袭数量明显增多(P<0.01)。相比对照siRNA组,Gadd45α-siRNA组滋养细胞HTR8/Svneo侵袭相关分子MMP-1、MMP-2、MMP-3及MMP-9的表达水平均显著升高(P<0.05)。结论Gadd45α高表达于PE胎盘组织,而通过阻滞滋养细胞HTR8/Svneo中Gadd45α的表达水平可诱导细胞侵袭,提高侵袭相关分子MMPs的表达量。

Objective To analyze the expression of growth retardation and DNA damage 45alpha(Gadd45α)protein and mRNA in placenta of preeclampsia(PE),and the effect of siRNA transfection on trophoblast invasion and invasion-related molecules.Methods In this retrospective study,40 cases of PE placental tissue were used as case group,and 30 cases of normal placental tissue were used as control group.The expression level of Gadd45αin the two groups was detected by immunoblotting,and the relative expression level of Gadd45αin the two groups was detected by real-time fluorescence quantitative PCR.Trophoblast HTR8/Svneo cells were cultured.The siRNA group transfected with negative control was the control siRNA group,and the siRNA group transfected with Gadd45αwas the Gadd45α-siRNA group.The number of invasive trophoblasts and the expression of matrix metalloproteinases(MMPs)were detected after transfection with siRNA of Gadd45α.Results Compared with the control group,the expression level of Gadd45αprotein in the case group was significantly increased(P<0.01).Compared with the control group,the relative expression level of Gadd45αgene in the case group was significantly increased(P<0.01).Compared with the control siRNA group,the expression level of Gadd45αin HTR8/Svneo trophoblasts in Gadd45α-siRNA group decreased significantly(P<0.01),and the relative expression level of Gadd45αwas also significantly decreased(P<0.01).With the increase of siRNA transfection time,the number of invasive cells in control siRNA group and Gadd45α-siRNA group increased significantly(P<0.05).Compared with control siRNA group,the number of HTR8/Svneo invasive cells in Gadd45α-siRNA group increased significantly(P<0.05).Compared with the control siRNA group,the expression levels of HTR8/Svneo invasion-related molecules MMP-1,MMP-2,MMP-3 and MMP-9 in the Gadd45α-siRNA group were significantly increased(P<0.01).Conclusion Gadd45αis highly expressed in the placenta of PE.Blocking the expression of Gadd45αin HTR8/Svneo can induce cell invasion and increase the expression of MMPs.