CircRNA-79530与TWIST形成正反馈回路调控滋养细胞EMT的机制

Mechanism of regulation of EMT of trophoblast cells via positive feedback loop by CircRNA-79530 and TWIST

目的探究CircRNA-79530与TWIST形成正反馈回路调控滋养细胞EMT的机制。方法筛选20例严重早发先兆子痫的患者和20例非严重早发先兆子痫的患者的胎盘样本。重组荧光素酶实验检测TWIST与CircRNA-79530靶标关系;RT-PCR检测TWIST和CircRNA-79530mRNA表达;Ed U染色法和CCK-8法测定细胞增殖;Annexin V-FITC检测细胞凋亡;蛋白印迹检测EMT蛋白表达;Transwell细胞迁移和侵袭实验检测细胞迁移侵袭能力;蛋白质免疫印迹法检测凋亡蛋白表达。结果重度先兆子痫组较非重度先兆子痫组TWIST、CircRNA-79530mRNA表达降低(P<0.05)。对照组较TWIST转染组细胞凋亡率升高,细胞活力降低,E-钙粘着蛋白、波形蛋白、β-连环蛋白、MMP9表达降低,细胞迁移和侵袭减少,caspase-3、Bax蛋白表达升高,Bcl-2蛋白表达降低(P<0.05)。结论TWIST直接与CircRNA-79530结合形成正反馈,促进滋养细胞的增殖、迁移侵袭。

Objective To explore the mechanism of regulation of EMT of trophoblast cells via positive feedback loop by CircRNA-79530 and TWIST.Methods Placental samples were screened from 20 women with severe early onset preeclampsia and 20 women without severe early onset preeclampsia.The expression of TWIST and circrna-79530 mRNA was detected by RT-PCR;cell proliferation by EdU staining and CCK-8;apoptosis by annexin V-FITC;EMT by Western blot;cell migration and invasion were detected by Transwell cell migration and invasion,and apoptotic factors by WB.Results The expression of TWIST and circrna-79530 m RNA in severe preeclampsia group was lower than that in non-severe preeclampsia group(P<0.05).The expression of E-cadherin,vimentin,β-Catenin,MMP9,cell migration and invasion,Caspase-3,Bax protein and bcl-2 protein in the control group were lower than those in TWIST group(P<0.05).Conclusion TWIST combines directly with circrna-79530 to form positive feedback,which promotes the proliferation,migration and invasion of trophoblasts.