PTEN regulation by miR-486-5p contributes to the amelioration of polycystic ovary syndrome.

Abstract

This research intended to identify the genes related to PCOS (Polycystic ovary syndrome) and verify the regulatory function of miR-486-5p as well as its target PTEN in granulosa cells (GCs). RT-qPCR was used to detect the expression of miR-486-5p in the serum, follicular fluid (FF), and granulosa cells (GC) of PCOS patients and normal subjects. ROC curve analysis indicated strong diagnostic performance. Bioinformatic analysis via miRDB and ENCORI databases predicted PTEN as a potential target of miR-486-5p, this prediction was validated through dual-luciferase reporter gene assays. Meanwhile, a series of functional assays were performed. Cellular proliferation capacity was quantitatively assessed using the CCK8 assay while flow cytometry was implied to determine cell apoptosis ratio. The secretion of pro-inflammatory mediators was quantitatively measured employing an ELISA kit. miR-486-5p was found to be reduced in serum from patients, as well as in patient FF and GCs. The enhanced expression of miR-486-5p strengthened the proliferation of GC and suppressing apoptotic activity, while concurrently attenuating pro-inflammatory cytokine secretion. Conversely, miR-486-5p inhibitor yielded opposing effects. Further investigation revealed that PTEN functioned as a negative regulatory factor of miR-486-5p. The increase of miR-486-5p caused a significant down-regulation of PTEN mRNA expression. Forced expression of PTEN reversed the cellular effects induced by miR-486-5p, including the enhanced proliferation rate, suppressed apoptosis, and attenuated inflammatory response. miR-486-5p can inhibit cell apoptosis and secretion of inflammatory factors by negatively regulating the expression of target gene PTEN, suggesting that miR-486-5p may be a potential target for PCOS.