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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.19.0290    [Accepted] Published online August 26, 2019.
MiR-126-3p inhibits apoptosis and promotes proliferation by targeting PIK3R2 in porcine ovarian granulosa cells
Xiaofeng Zhou1  , Yingting He1  , Yao Jiang1  , Bo He1  , Xi Deng1  , Zhe Zhang1  , Xiaolong Yuan1  , Jiaqi Li1,* 
Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breeding, National Engineering Research Centre for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, Guangdong, China
Correspondence:  Jiaqi Li, Tel: +86-0-20-85283519, Fax: +86-0-20-85283519, Email: jqli@scau.edu.cn
Received: 7 April 2019   • Revised: 15 June 2019   • Accepted: 14 August 2019
Abstract
Objective
s: Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored.
Methods
In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2’-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative PCR and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression.
Results
We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1).
Conclusion
MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1 and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.
Keywords: Ovarian Granulosa Cells; miR-126-3p; PIK3R2; Cell Apoptosis; Cell Proliferation
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