MicroRNA-664a-3p inhibits the proliferation of ovarian granulosa cells in polycystic ovary syndrome and promotes apoptosis by targeting BCL2A1

Min He; Ganghong Mao; Yungai Xiang; Pengfen Li; Yuanyuan Wu; Dongmei Zhao; Tan Li

doi:10.21037/atm-21-1614

MicroRNA-664a-3p inhibits the proliferation of ovarian granulosa cells in polycystic ovary syndrome and promotes apoptosis by targeting BCL2A1

Ann Transl Med. 2021 May;9(10):852. doi: 10.21037/atm-21-1614.

Authors

Min He¹, Ganghong Mao¹, Yungai Xiang¹, Pengfen Li¹, Yuanyuan Wu¹, Dongmei Zhao¹, Tan Li¹

Affiliation

¹ Department of Reproductive Medicine, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, China.

Abstract

Background: To investigate whether micro ribonucleic acid-664a-3p (miR-664a-3p) targeting BCL2A1 affects the proliferation and apoptosis of ovarian granulosa cells.

Methods: Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-664a-3p in granulosa cells, granular tumor cell lines (KGN), and normal ovarian epithelial cell lines (IOSE80) in the polycystic ovary syndrome (PCOS) group and the control group. After overexpressing miR-664a-3p or inhibiting its expression in KGN cells, qRT-PCR and Western blotting were used to detect the messenger RNA (mRNA) and protein levels of related genes. At the same time, a cell counting kit-8 (CCK-8) and flow cytometer were used to detect cell proliferation and apoptosis. The TargetScan website was used to predict the potential binding sites of miR-664a-3p and B-cell lymphoma 2-related protein A1 (BCL2A1), which was further verified by qRT-PCR, Western blotting, and the luciferase reporter gene method.

Results: The expression of miR-664a-3p was significantly decreased in both PCOS tissues and KGN cells (both P<0.05), and the overexpression of miR-664a-3p inhibited the proliferation of KGN cells and induced their apoptosis. Moreover, our results confirmed that miR-664a-3p directly targets BCL2A1 (P<0.05), and the inhibitory effect of miR-664a-3p on KGN cells was reversed by BCL2A1 overexpression (both P<0.05). The up-regulation of BCL2A1 promotes cell proliferation and reduces cell apoptosis by the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway (both P<0.05).

Conclusions: The up-regulation of miR-664a-3p inhibits the proliferation of KGN cells and increases apoptosis by down-regulating the expression of BCL2A1 and blocking the MAPK/ERK signaling pathway.

Keywords: B-cell lymphoma 2-related protein A1 (BCL2A1); Polycystic ovary syndrome (PCOS); apoptosis; micro ribonucleic acid-664a-3p (miR-664a-3p); ovarian granulosa cells; proliferation.