摘要
目的探讨miR-191-5p靶向调节AT富集序列特异性结合蛋白1(SATB1),对子痫前期(pre-eclampsia,PE)滋养层细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)及细胞迁移的影响.方法收集滋养层细胞系HTR-8/SVneo及人绒毛膜癌细胞系JEG-3、JAR.分组转染后qRT-PCR及Western blot检测细胞中miR-191-5p及SATB1的表达;Transwell检测细胞迁移;qRT-PCR及Western blot检测迁移侵袭相关因子MMP-9、MMP-2的表达;Western blot检测上皮间质转化相关因子的蛋白表达.结果miR-191-5p在HTR-8/SVneo细胞系中高表达,SATB1低表达.Western blot检测发现过表达miR-191-5p后,SATB1蛋白水平表达下调,抑制miR-191-5p的表达后表达上调(均P<0.05).过表达miR-191-5p,细胞增殖、迁移能力降低,MMP-2及MMP-9表达下调,抑制细胞EMT的发生,抑制miR-191-5p表达则结果相反.而抑制SATB1表达后,与过表达miR-191-5p结果一致,过表达SATB1后结果与抑制miR-191-5p结果一致.结论MiR-191-5p可靶向调节SATB1,降低细胞迁移能力,并抑制HTR-8/SVneo细胞EMT参与PE的发生.
Objective To explore the effect of miR⁃191⁃5p on epithelial⁃mesenchymal transition(EMT)and migration of trophoblastic cells in pre⁃eclampsia(PE)through targeting SATB1.Methods Trophoblast cell line HTR⁃8/Svneo and choriocarcinoma cell line JEG⁃3、JAR were collected.qRT⁃PCR and Western blot were utilized to detect miR⁃191⁃5p and SATB1 expression after cell grouping and transfection.Transwell was performed to detect cell migration.Expression of migration and invasion related factors MMP⁃9、MMP⁃2 was quantified through qRT⁃PCR and Western blot.At the same time,protein expression of EMT related factors was detected by Western blot.Results MiR⁃191⁃5p expression was up⁃regulated while SATB1 expression was suppressed in HTR⁃8/Svneo cell line.Western blot results showed that,SATB1 protein expression was decreased when miR⁃191⁃5p was overexpressed,SATB1 expression was increased when miR⁃191⁃5p expression was inhibited(all P<0.05).It was found that cell proliferation and migration,the expression of MMP⁃2,MMP⁃9 were decreased,EMT in cells was also inhibited when miR⁃191⁃5p was overexpressed,However,the results contrast with above outcomes when miR⁃191⁃5p was inhibited.Results obtained from si⁃SATB1 group and pcDNA⁃SATB1 group were consistent with that in mimics miR⁃191⁃5p group.Conclusion MiR⁃191⁃5p influence the formation of PE by downregulating the migration and EMT of HTR⁃8/Svneo cells through targeting SATB1.
作者
周俏苗
陈秋霞
黄海燕
许晶
肖美芳
龚护民
吴栋才
ZHOU Qiaomiao;CHEN Qiuxia;HUANG Haiyan;XU Jing;XIAO Meifang;GONG Humin;WU Dongcai(Department of Obstetrics and Gynecology,Hainan Women and Children Medical Center,Haikou 570000,China;不详)
出处
《实用医学杂志》
CAS
北大核心
2020年第12期1584-1589,共6页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号:81760275)。
