摘要
目的 探讨LncRNA PSMA3-AS1对子宫内膜癌细胞增殖、迁移、侵袭的影响及分子机制.方法 收集内蒙古自治区人民医院38例子宫内膜癌患者癌组织及癌旁组织,培养正常子宫内膜上皮细胞hEEC及子宫内膜癌细胞系Ishikawa、HEC-1A、AN3CA,RT-qPCR检测RNA水平;将Ishikawa细胞分为NC组、si-LncRNAPSMA3-ASI 组、si-NC组、miR-329-3p组、miR-NC 组、anti-miR-329-3p+si-LncRNAPSMA3-AS1 组和anti-miR-329-3p+si-NC组;Transwell检测细胞迁移和侵袭;Western blot法检测蛋白表达;CCK-8法检测细胞活性;双荧光素酶报告实验分析荧光素酶活性.结果 与癌旁组织比较,子宫内膜癌组织中miR-329-3p表达水平降低,LncRNAPSMA3-AS1表达水平升高(P＜0.05).与hEEC细胞比较,子宫内膜癌细胞系Ishikawa、HEC-1A、AN3CA 中 miR-329-3p 表达水平降低,LncRNA PSMA3-AS1 表达水平升高(P＜0.05).LncRNA PSMA3-AS1 低表达或miR-329-3p高表达,CyclinD1、MMP2、MMP9蛋白表达水平降低,Ishikawa细胞活性降低,Ishikawa细胞迁移、侵袭数减少(P＜0.05).LncRNA PSMA3-AS1靶向调控miR-329-3p的表达;抑制miR-329-3p表达可以逆转LncRNAPSMA3-AS1低表达对Ishikawa细胞的影响.结论 LncRNAPSMA3-AS1通过靶向抑制miR-329-3p的表达促进子宫内膜癌Ishikawa细胞恶性表型的发展.
Abstract
Objective To explore the effect of LncRNA PSMA3-AS1 on the proliferation,migration and invasion of endometrial cancer cells and its molecular mechanism.Methods We collected tissue samples from 38 endometrial cancer patients,including cancer tissues and adjacent tissues in our hospital.We cultured normal endometrial epithelial cells(hEEC)and endometrial cancer cell lines(Ishikawa,HEC-1A,AN3CA)and employed RT-qPCR to detect RNA level;Ishikawa cells were divided into NC group,si-LncRNA PSMA3-AS1 group,si-NC group,miR-329-3p group,miR-NC group,anti-miR-329-3p+si-LncRNA PSMA3-AS1 group,anti-miR-329-3p+si-NC group;We assessed cell invasive and migratory migration capacities using transwell assays;measured protein contents through Western blot;evaluated cell activity using the cell counting kit 8(CCK-8),and conducteddual luciferase reporter experiment to analyze luciferase activities.Results MiR-329-3p was decreased,but LncRNA PSMA3-AS1 was increased in endometrial cancer tissues in comparison with adjacent tissues(P＜0.05).Compared with hEEC cells,the expression level of miR-329-3p in endometrial cancer cell lines Ishikawa,HEC-1A,AN3CA was decreased,and the expression level of LncRNA PSMA3-AS1 was increased(P＜0.05).LncRNA PSMA3-AS1 expression was low or miR-329-3p expression was high,the protein expression levels of CyclinDl,MMP2 and MMP9 were decreased,the activity of Ishikawa cells was decreased,and the number of migration and invasion of Ishikawa cells was decreased(P＜0.05).LncRNA PSMA3-AS1 was found to target miR-329-3p,and inhibiting the expression of miR-329-3p reversed the effects induced by low expression of LncRNA PSMA3-AS1 in Ishikawa cells.Conclusion LncRNA PSMA3-AS1 promotes the malignant phenotype of Ishikawa endometrial cancer cells by targeting the inhibition of miR-329-3p expression.
基金项目
内蒙古自治区自然科学基金(2019MS08098)
NETL
NSTL
维普
万方数据