Department of Obstetrics and Gynecology Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Department of Obstetrics and Gynecology Surgery, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450014, P.R. China.
Mol Med Rep. 2017 Oct;16(4):5706-5712. doi: 10.3892/mmr.2017.7270. Epub 2017 Aug 17.
MicroRNAs (miRNAs) are widely involved in regulation of cellular processes of polycystic ovary syndrome (PCOS). However, the function of miR‑320a in PCOS remains unclear. The present study aimed to explore the effect of miR‑320a on PCOS cell proliferation and apoptosis following treatment with insulin, and to clarify the underlying mechanism. PCOS tissues and corresponding normal tissues were collected from 16 female patients with PCOS. KGN cells were pre‑treated with insulin, and KGN cells were transfected with ASO‑miR‑320a, miR‑320a mimics and polycomb group ring finger 1 (PCGF1) overexpression plasmids. Expressions of miR‑320a and PCGF1 were detected using the reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Dual‑Luciferase reporter assays were performed to investigate the target gene of miR‑320a. MTS, colony formation and flow cytometry assays were performed to determine cell viability, colony formation, and apoptosis, respectively. Furthermore, mRNA and protein expression levels of B‑cell lymphoma 2 apoptosis regulator (Bcl‑2) and Bcl‑2 associated protein X apoptosis regulator (Bax) were examined using RT‑qPCR and western blotting. The results demonstrated that miR‑320a expression was significantly increased in PCOS tissues compared with normal tissues. Moreover, miR‑320a was upregulated in insulin‑induced cells in a dose‑dependent manner. Inhibition of miR‑320a suppressed insulin‑induced cell viability and colony formation, and promoted apoptosis. Luciferase reporter assays demonstrated that PCGF1 was a target of miR‑320a. Additionally, PCGF1 overexpression inhibited cell viability and colony formation and promoted apoptosis. Additionally, the mRNA and protein levels of Bcl‑2 were inhibited by miR‑320a suppression and PCGF1 overexpression, while Bax expression was promoted by them in insulin‑induced cells. The results of the present study demonstrated that miR‑320a inhibition decreased insulin‑induced KGN cell proliferation and apoptosis by targeting PCGF1. These data indicated that miR‑320a may serve as a potential diagnostic biomarker for PCOS.
微小 RNA（miRNA）广泛参与多囊卵巢综合征（PCOS）的细胞过程调节。然而，miR-320a 在 PCOS 中的功能仍不清楚。本研究旨在探讨 miR-320a 对胰岛素处理后 PCOS 细胞增殖和凋亡的影响，并阐明其潜在机制。收集 16 例 PCOS 女性患者的 PCOS 组织和相应的正常组织。用胰岛素预处理 KGN 细胞，并用 ASO-miR-320a、miR-320a 模拟物和多梳组蛋白 1（PCGF1）过表达质粒转染 KGN 细胞。采用逆转录-定量聚合酶链反应（RT-qPCR）检测 miR-320a 和 PCGF1 的表达。进行双荧光素酶报告基因检测以研究 miR-320a 的靶基因。采用 MTS、集落形成和流式细胞术分别检测细胞活力、集落形成和凋亡。此外，采用 RT-qPCR 和 Western blot 检测 B 细胞淋巴瘤 2 凋亡调节因子（Bcl-2）和 Bcl-2 相关蛋白 X 凋亡调节因子（Bax）的 mRNA 和蛋白表达水平。结果表明，与正常组织相比，PCOS 组织中 miR-320a 的表达显著增加。此外，miR-320a 在胰岛素诱导的细胞中呈剂量依赖性上调。抑制 miR-320a 可抑制胰岛素诱导的细胞活力和集落形成，并促进凋亡。荧光素酶报告基因检测表明，PCGF1 是 miR-320a 的靶基因。此外，过表达 PCGF1 可抑制细胞活力和集落形成并促进凋亡。此外，抑制 miR-320a 和过表达 PCGF1 可抑制胰岛素诱导的细胞中 Bcl-2 的 mRNA 和蛋白表达水平，同时促进 Bax 的表达。本研究结果表明，miR-320a 通过靶向 PCGF1 抑制胰岛素诱导的 KGN 细胞增殖和凋亡。这些数据表明，miR-320a 可能作为 PCOS 的潜在诊断生物标志物。
