MicroRNA‑320a inhibition decreases insulin‑induced KGN cell proliferation and apoptosis by targeting PCGF1

Mol Med Rep. 2017 Oct;16(4):5706-5712. doi: 10.3892/mmr.2017.7270. Epub 2017 Aug 17.

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.

MeSH terms

  • 3' Untranslated Regions
  • Apoptosis / genetics*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Female
  • Gene Expression
  • Gene Expression Regulation
  • Genes, Reporter
  • Granulosa Cells / metabolism*
  • Humans
  • Insulins / metabolism*
  • Insulins / pharmacology
  • MicroRNAs / genetics*
  • Polycomb Repressive Complex 1 / genetics*
  • Polycystic Ovary Syndrome / genetics
  • Polycystic Ovary Syndrome / metabolism
  • Polycystic Ovary Syndrome / pathology
  • RNA Interference

Substances

  • 3' Untranslated Regions
  • Insulins
  • MIRN320 microRNA, human
  • MicroRNAs
  • PCGF1 protein, human
  • Polycomb Repressive Complex 1