摘要
目的:研究锌指转录因子Snail高表达对子痫前期(PE)大鼠胎盘滋养细胞侵袭能力的影响,并探讨其相关机制。方法:建立大鼠PE模型,作为模型组,取健康大鼠作为对照组。从PE大鼠绒毛膜滋养层组织分离和培养原代滋养细胞。取对数生长期滋养细胞,分为Neo组(转染Snail阴性对照质粒pL-tdTomato-Neo)和mSnail组(转染Snail过表达质粒pL-tdTomato-mSnail),以不作处理的细胞为空白对照组。采用实时荧光定量PCR(RT-qPCR)和Western blotting法检测对照组和模型组大鼠胎盘绒毛膜滋养层组织及空白对照组、Neo组和mSnail组细胞中Snail mRNA和蛋白表达水平,Transwell小室实验检测各组细胞侵袭能力,Western blotting法检测各组细胞中上皮性钙黏蛋白(E-cadherin)、神经性钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平。结果:与对照组比较,模型组大鼠胎盘绒毛膜滋养层组织中Snail mRNA和蛋白表达均明显降低(P<0.05);荧光显微镜下观察,Neo组和Snail组细胞转染效率均>80%。与空白对照组和Neo组比较,mSnail组细胞中Snail mRNA和蛋白表达水平明显升高(P<0.05),每个视野平均穿过微孔的细胞数明显增加(P<0.05),E-cadherin蛋白表达水平明显降低(P<0.05),N-cadherin和Vimentin蛋白表达水平明显升高(P<0.05)。结论:Snail高表达可增强PE大鼠胎盘滋养细胞的侵袭能力,其机制可能与Snail促进胎盘滋养细胞上皮-间质转化过程有关。
Objective:To study the effect of high expression of zinc finger transcription factor Snail on the invasion capacity of placental trophoblasts in the preeclampsia(PE)rats,and to explore the related mechanisms. Methods:The PE rat models were established and used as model group;the healthy rats were selected and used as control group. The primary trophoblasts were isolated and cultured from the chorionic trophoblast tissue of the PE rats. The trophoblasts in the logarithmic growth phase were selected as Neo group(transfected with pL-tdTomato-Neo plasmid) and mSnail group(transfected with pLtdTomato-mSnail plasmid),and the untreated cells were used as blank control group. Real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting methods were used to detect the expression levels of Snail mRNA and protein in chorionic trophoblast tissue of the rats in control group and model group and in the cells in blank control group,Neo group and mSnail group. Transwell chamber experiment was used to determine the cell invasion abilities in blank control group,Neo group and mSnail group. Western blotting method was used to detect the expression levels of epithelial cadherin(Ecadherin),neurogenic cadherin(N-cadherin)and Vimentin proteins in the cells. Results:Compared with control group,the expression levels of Snail mRNA and protein in the placenta chorionic trophoblast tissue of the rats in model group were decreased(P<0. 05);under fluorescence microscope,the transfection efficiencies in Neo group and mSnail group were both >80%. Compared with blank control group and Neo group,the expression levels of Snail mRNA and protein in the cells in mSnail group were increased(P<0. 05),the average number of cells passing through the micropore in each field of view was increased(P<0. 05),the expression level of E-cadherin protein was decreased(P<0. 05),and the expression levels of Ncadherin and Vimentin proteins were increased(P<0. 05). Conclusion High expression of Snail can enhance the invasion ability of placental trophoblasts
作者
张长存
张爱萍
李玉琴
ZHANG Changcun;ZHANG Aiping;LI Yuqin(Department of Obstetrics,Qinghai Provincial People’s Hospital,Xining 810000,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2021年第4期911-918,共8页
Journal of Jilin University：Medicine Edition
基金
青海省科技厅科研项目(9632018Y0152)。
