Fzr1基因沉默促进滋养层细胞的侵袭与增殖

Promotion of the invasion and proliferation of trophoblast cells by silencing Fzr1

目的观察Fzr1蛋白在胎盘发育过程中时空性表达变化及其对滋养层细胞侵袭与增殖的影响。方法免疫印迹和免疫组化法观察Fzr1蛋白的表达量和表达部位,通过RNA干扰技术、Transwell实验和MTS法检测Fzr1对滋养层细胞侵袭和增殖。结果 1)在妊娠早期,Fzr1蛋白强烈表达于绒毛的细胞滋养层和合体滋养层细胞,阳性信号主要位于胞质。在妊娠晚期,Fzr1蛋白主要表达于合体滋养层细胞,且表达量明显下降,阳性信号主要定位于胞核。与妊娠早期相比,妊娠晚期Fzr1蛋白下降4.88倍(P<0.01)。2)与空白对照组相比,Fzr1干扰组,JAR细胞的侵袭能力明显增强(P<0.01),而无义对照,没有明显变化。3)与空白对照组相比,Fzr1干扰组,JAR细胞的增殖能力明显增强(P<0.01),而无义对照,没有明显变化。结论 Fzr1蛋白在正常妊娠胎盘呈现时空性的表达,Fzr1基因沉默能促进滋养层细胞的侵袭和增殖能力。

Objective To explore the spatial and temporal expression of Fzr1 protein during the development of placenta, and observe the effect of Fzr1 on the invasion and proliferation of trophoblast cells. Methods To observe the expression level and localization of Fzr1 protein using Western blot and immunohistochemical methods, and to detect the effect of Fzr1 on trophoblast cell invasion and proliferation by RNA interference technology, Transwell and MTS methods. Results 1 ） Fzr1 protein was significantly expressed in the cytotrophoblast and syncy-tiotrophoblast cells of villi during early pregnancy. Positive signals were mainly located in the cytoplasm. In the third trimester of pregnancy, Fzr1 protein was expressed in the syncytiotrophoblast, and expression level significantly decreased. Positive signal was located in the nucleus. Western blot studies showed that Fzr1 protein levels in the third trimester decreased by 4. 88 times as compared with early pregnancy （ P 〈 0.01 ）. 2 ） Fzr1 siRNA significantly enhanced the invasion of JAR cells compared with blank control （ P 〈 0.01 ） , but scrambled siRNA had no significantly change. 3 ）Fzr1 siRNA significantly enhanced the proliferation of JAR cells compared with blank control （P 〈 0.01 ） , but scrambled siRNA had no significantly change. Conclusions Fzr1 protein is temporally and spatially expressed at the placenta of normal pregnancy, and Fzr1 gene silencing may promote the invasion and proliferation of trophoblast cell.