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Published on: May 21, 2019
MicroRNA‑320a inhibition decreases insulin‑induced KGN cell proliferation and apoptosis by targeting PCGF1
Lei Yang1, Yan Li1, Xiaojing Wang2
1Department of Obstetrics and Gynecology Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Abstract:
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.
Insights
MicroRNA-320a (miR-320a) is elevated in polycystic ovary syndrome (PCOS) and promotes PCOS cell proliferation by targeting PCGF1. Inhibiting miR-320a reduces proliferation and enhances apoptosis in PCOS cells, suggesting its potential as a diagnostic biomarker.
Area of Science:
- Endocrinology
- Molecular Biology
- Cell Biology
Background:
- MicroRNAs (miRNAs) play a role in polycystic ovary syndrome (PCOS) cellular processes.
- The specific function of miR-320a in PCOS pathogenesis is not well understood.
Purpose of the Study:
- To investigate the effect of miR-320a on PCOS cell proliferation and apoptosis.
- To elucidate the underlying molecular mechanism involving insulin treatment and target gene identification.
Main Methods:
- Analysis of miR-320a and PCGF1 expression in PCOS tissues and insulin-treated KGN cells.
- Luciferase reporter assays to confirm miR-320a targeting of PCGF1.
- Cell viability, colony formation, apoptosis assays, and Western blotting for apoptosis-related proteins (Bcl-2, Bax).
Main Results:
- miR-320a expression was significantly increased in PCOS tissues and upregulated by insulin in KGN cells.
- Inhibition of miR-320a suppressed cell viability and colony formation, and promoted apoptosis.
- PCGF1 was identified as a direct target of miR-320a; PCGF1 overexpression mimicked the effects of miR-320a inhibition.
- miR-320a inhibition and PCGF1 overexpression altered Bcl-2 and Bax expression, impacting apoptosis.
Conclusions:
- miR-320a inhibition decreases insulin-induced KGN cell proliferation and apoptosis by targeting PCGF1.
- miR-320a may serve as a potential diagnostic biomarker for PCOS.
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