LncRNA WDR86-AS1调节FOXO3a对滋养细胞增殖和侵袭能力的影响

LncRNA WDR86-AS1 affected the trophoblast proliferation and invasion by regulating FOXO3a

目的:探究LncRNA WDR86-AS1对叉头框转录因子3a(FOXO3a)与胎盘组织和人早孕绒毛外滋养细胞HTR8/SVneo细胞增殖和侵袭能力的影响及其可能参与子痫前期(PE)发生发展的分子机制.方法:选择子痫前期(PE)孕妇的胎盘组织25例为PE组,同期健康孕妇的胎盘组织25例为对照组,采用免疫组化法、蛋白印迹法及实时荧光定量PCR技术检测两组孕妇胎盘组织中LncRNA WDR86-AS1和FOXO3a的表达水平,并分析二者表达的相关性;沉默HTR8/SVneo细胞中LncRNA WDR86-AS1或FOXO3a的表达,检测FOXO3a的表达,并采用CCK-8和Transwell检测对滋养细胞增殖和侵袭能力的影响.结果:PE组中LncRNA WDR86-AS1 mRNA、FOXO3a mRNA和蛋白表达均高于对照组(P<0.05);FOXO3a在胎盘组织滋养细胞的胞质和胞核均有表达且PE组的表达高于对照组(P<0.05);PE组中LncRNA WDR86-AS1和FOXO3a表达呈正相关(r=0.54,P<0.05);在HTR8/SVneo细胞中沉默LncRNA WDR86-AS1,FOXO3a mRNA和蛋白表达水平下降(P<0.05),细胞的增殖和侵袭能力增加(P<0.05);在HTR8/SVneo细胞中沉默FOXO3a,FOXO3a mRNA和蛋白表达水平下降(P<0.05),细胞的增殖和侵袭能力增加(P<0.05).结论:LncRNA WDR86-AS1可能通过调节FOXO3a的表达影响滋养细胞增殖和侵袭能力,参与PE的发生发展.

Objective:To investigate the influence of LncRNA WDR86-AS1 to FOXO3a and the placental tissues and HTR8/SVneo cells trophoblast proliferation and invasion,which could involve the molecular mechanisms in preeclampsia(PE).Methods:Twenty-five preeclampsia placental tissues as the PE group and 25 healthy pregnancy placental tissues as the control group were selected to detected the expression of LncRNA WDR86-AS1 and FOXO3a by immunohistochemistry、Western blotting and qRT-PCR.And the correlation of mRNA expression between LncRNA WDR86-AS1 and FOXO3a was detected in preeclampsia placental tissues.After transfecting LncRNA WDR86-AS1 siRNA into the HTR8/SVneo cells,the mRNA and protein expression of FOXO3a were detected through qRT-PCR and Western blotting,the proliferation of HTR8/SVneo cells were determined through CCK-8,and the invasion of HTR8/SVneo cells were detected by transwell assay.Results:The mRNA and protein expression of LncRNA WDR86-AS1 and FOXO3a were increased in PE group(P<0.05)and the mRNA expression of LncRNA WDR86-AS1 was positive correlated with FOXO3a in PE group(r=0.54,P<0.05).The expression of FOXO3a was elevated in cytoplasm and nucleus of trophoblast cells in PE group(P<0.05).After transfecting LncRNA WDR86-AS1 siRNA into the HTR8/SVneo cells,the mRNA and protein expression of FOXO3a were suppressed,and the proliferation and invasion in HTR8/SVneo cells were increased(P<0.05).After transfecting FOXO3a siRNA into the HTR8/SVneo cells,the mRNA and protein expression of FOXO3a were suppressed,and the proliferation and invasion in HTR8/SVneo cells were increased(P<0.05).Conclusion:The LncRNA WDR86-AS1 could affect the trophoblast proliferation and invasion by regulating the expression of FOXO3a,which may be involved in preeclampsia.