Department of Reproductive Medicine, Liaocheng People's Hospital, NO.67, Dongchang West Road, Shandong Province, 252000, Liaocheng City, P. R. China.
College of Bioengineering, Wuhu Institute of Technology, NO.201, Wenjin West Road, Wuhu City, Anhui Province, 241103, P. R. China.
J Ovarian Res. 2021 Jan 23;14(1):19. doi: 10.1186/s13048-020-00742-4.
Currently, several non-coding RNAs (ncRNAs) were distinguished in polycystic ovarian syndrome (PCOS). This present study aims to explore the potential function of lncRNA HOTAIRM1/miR-433-5p/PIK3CD in ovarian granulosa cells.
We analyzed the expression profiles of HOTAIRM1, miR-433-5p and PIK3CD in PCOS samples by enquiring GEO database. GSEA was applied to enrich the pathways related to PCOS. The target association between HOTAIRM1 and miR-433-5p or the binding association between miR-433-5p and PIK3CD were assessed by online prediction tools and a dual luciferase reporter assay. qPCR and western blotting assays were used to detect PIK3CD expression after HOTAIRM1 and miR-433-5p treatment. The proliferation and apoptosis of ovarian granulosa cells were estimated by cell counting kit-8 and flow cytometry assays, respectively.
The expression profiles of HOTAIRM1 and PIK3CD were increased, whereas miR-433-5p was decreased in PCOS tissues. PIK3CD expression was positively regulated by HOTAIRM1 and negatively modulated by miR-433-5p. Overexpression of HOTAIRM1 reduced the proliferative ability and increased the apoptotic ability of granulosa cells, whereas upregulation of miR-433-5p or downregulation of PIK3CD reversed the effects of HOTAIRM1 on granulosa cells. Moreover, overexpression of miR-433-5 displayed a results with increasing proliferative ability and decreasing apoptotic ability, but upregulation of PIK3CD eliminated the function of miR-433-5p on granulosa cells.
Our findings illustrated that HOTAIRM1 could sponge miR-433-5p to promote PIK3CD expression, thereby regulating the growth and apoptosis of granulose cells in PCOS.
目前,多囊卵巢综合征(PCOS)中已区分出几种非编码 RNA(ncRNA)。本研究旨在探讨长链非编码 RNA HOTAIRM1/miR-433-5p/PIK3CD 在卵巢颗粒细胞中的潜在功能。
我们通过查询 GEO 数据库分析了 PCOS 样本中 HOTAIRM1、miR-433-5p 和 PIK3CD 的表达谱。应用 GSEA 富集与 PCOS 相关的通路。通过在线预测工具和双荧光素酶报告基因实验评估 HOTAIRM1 与 miR-433-5p 或 miR-433-5p 与 PIK3CD 之间的靶标关联。qPCR 和 Western blot 检测 HOTAIRM1 和 miR-433-5p 处理后 PIK3CD 的表达。通过细胞计数试剂盒-8 和流式细胞术分别评估卵巢颗粒细胞的增殖和凋亡。
PCOS 组织中 HOTAIRM1 和 PIK3CD 的表达水平升高,而 miR-433-5p 的表达水平降低。PIK3CD 的表达受 HOTAIRM1 正向调节,受 miR-433-5p 负向调节。HOTAIRM1 过表达降低了颗粒细胞的增殖能力并增加了其凋亡能力,而 miR-433-5p 的上调或 PIK3CD 的下调则逆转了 HOTAIRM1 对颗粒细胞的作用。此外,miR-433-5p 的过表达显示出增加增殖能力和降低凋亡能力的结果,但 PIK3CD 的上调消除了 miR-433-5p 对颗粒细胞的功能。
我们的研究结果表明,HOTAIRM1 可以通过海绵吸附 miR-433-5p 来促进 PIK3CD 的表达,从而调节 PCOS 中颗粒细胞的生长和凋亡。