lncRNA TP73-AS1通过调控miR-128-3p对多囊卵巢综合征卵巢颗粒细胞凋亡的影响
Effect of lncRNA TP73-AS1 on ovarian granulosa cell apoptosis in polycystic ovary syndrome by regulating miR-128-3p
孟睿 1张雯1
作者信息
- 1. 湖北省大冶市人民医院妇产科,湖北大冶 435199
- 折叠
摘要
目的 探讨lncRNA TP73-AS1在多囊卵巢综合征中的作用及潜在机制.方法 体外培养人卵巢颗粒细胞系 KGN 细胞,将 Vector、pcDNA-TP73-AS1、NC-siRNA、TP73-AS1-siRNA、TP73-AS1-siRNA+NC inhibitor、TP73-AS1-siRNA+miR-128-3p inhibitor 分别转染至 KGN 细胞,实时荧光定量 PCR(RT-qPCR)实验检测 lncRNA TP73-AS1、miR-128-3p的表达水平;MTT实验检测细胞增殖能力;流式细胞术检测细胞凋亡率;生物信息学软件和双荧光素酶报告基因实验预测和验证lncRNA TP73-AS1和miR-128-3p之间的靶向关系.结果 细胞转染pcDNA-TP73-AS1后,lncRNA TP73-AS1表达上调,细胞增殖能力升高,细胞凋亡率降低(P＜0.05).细胞转染TP73-AS1-siRNA后,lncRNA TP73-AS1表达下调,细胞增殖能力降低,凋亡率升高(P＜0.05).双荧光素酶报告基因实验结果显示,miR-128-3p 是 lncRNA TP73-AS1 的靶基因.TP73-AS1-siRNA 组细胞中 miR-128-3p 的表达较 NC-siRNA 组显著升高(P＜0.05);pcDNA-TP73-AS1 组细胞中 miR-128-3p 的表达较 Vector 组显著降低(P＜0.05).miR-128-3p in-hibitor处理可显著逆转TP73-AS1-siRNA对细胞增殖和凋亡的调控作用.结论 敲低lncRNA TP73-AS1可能通过靶向miR-128-3p抑制卵巢颗粒细胞增殖,促进细胞凋亡.
Abstract
Objective To investigate the role of lncRNA TP73-AS1 in polycystic ovary syndrome and its potential mechanism.Methods Human ovarian granulosa cell line KGN cells were cultured in vitro,and Vector,pcDNA-TP73-AS1,NC-siRNA,TP73-AS 1-siRNA,TP73-AS 1-siRNA+NC inhibitor,TP73-AS1-siRNA+miR-128-3p inhibitor were transfected into KGN cells,respectively.Real-time quantitative PCR(RT-qPCR)experiments were used to detect the expression levels of lncRNA TP73-AS1 and miR-128-3p.MTT assay was used to detect cell proliferation.The apoptosis rate was detected by flow cytometry.Bioinformatics software and dual luciferase reporter assay were used to predict and verify the targeting relationship between lncRNA TP73-AS1 and miR-128-3p.Results After cells were transfected with pcDNA-TP73-AS1,the expression of lncRNA TP73-AS1 was up-regulated,the cell proliferation ability was increased,and the apoptosis rate was decreased(P＜0.05).After cells were transfected with TP73-AS 1-siRNA,the expression of lncRNA TP73-AS1 was down-regulated,the cell proliferation ability was decreased,and the apoptosis rate was increased(P＜0.05).Dual luciferase reporter assay showed that miR-128-3p was the target gene of lncRNA TP73-AS1.The expression of miR-128-3p in TP73-AS1-siRNA group was significantly higher than that in NC-siRNA group(P＜0.05).The expression of miR-128-3p in pcDNA-TP73-AS1 group was significantly lower than that in Vector group(P＜0.05).miR-128-3p inhibitor treatment could significantly reverse the regulatory effects of TP73-AS1-siRNA on cell proliferation and apoptosis.Conclusion Knockdown of lncRNA TP73-AS1 could inhibit ovarian granulosa cell proliferation and promote apoptosis by targeting miR-128-3p.
关键词
多囊卵巢综合征/lncRNA/TP73-AS1/miR-128-3p/卵巢颗粒细胞/凋亡Key words
polycystic ovary syndrome/lncRNA TP73-AS1/miR-128-3p/ovarian granulosa cells/apoptosis引用本文复制引用
基金项目
湖北省卫生健康委联合基金立项项目(第一批)(2019)(WJ2019H190)
出版年
2024
NETL
NSTL
万方数据