miR‐221‐3p regulates apoptosis of ovarian granulosa cells via targeting FOXO1 in older women with diminished ovarian reserve (DOR)

Abstract

In our earlier study, we showed that the expression of microRNA‐221‐3p (miR‐221‐3p) was significantly lower in women of advanced age with diminished ovarian reserve (DOR) compared with young women with normal ovarian reserve (NOR). Therefore, in this study, we aimed to explore how miR‐221‐3p regulates apoptosis of granulosa cells and the pathogenesis of DOR. Bioinformatics prediction and dual‐luciferase reporter assay were conducted to identify the target gene of miR‐221‐3p. miR‐221‐3p expression was manipulated by transfecting KGN cells with miR‐221‐3p mimics, inhibitor, and negative control. Following transfection, apoptosis of granulosa cells was determined by flow cytometry, and the expression of the target gene was measured by quantitative real‐time polymerase chain reaction (qRT‐PCR) and western blot analysis (WB). In addition, the expression of the target gene in granulosa cells of DOR patients and NOR patients was measured. miR‐221‐3p were found to directly bind the 3ʹ untranslated region of Forkhead box O1 (FOXO1). Transfection with miR‐221‐3p mimics significantly decreased the apoptosis rate of KGN cells compared with transfection with miR‐221‐3p inhibitors. The expression level of miR‐221‐3p was negatively correlated with the messenger RNA and protein levels of the FOXO1 gene. Besides, FOXO1 expression was upregulated in DOR patients. In conclusion, these results provide evidence that downregulation of miR‐221‐3p expression promotes apoptosis of granulosa cells by upregulating FOXO1 expression, thus serving an important role in DOR pathogenesis.

Role of microRNAs in response to cadmium chloride in pancreatic ductal adenocarcinoma

“…miR-221 regulates a target, Dickkopf 2, subsequently inhibiting the activation of the Wnt/β-catenin pathways (Chen et al 2020 ). An inverse relationship between miR-221 and FOXO1 (downregulation of miR-221/upregulation of FOXO1) has been reported in ovarian cells in menopause (Wei et al 2021 ). Overexpression of miR-221 can directly increase the growth and proliferation of PDAC cells by downregulating PTEN expression and upregulating phospho-Akt expression (Yang et al 2016 ).…”

Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

“…Our experimental results revealed that high-dose administration of RD inhibited the damage caused by CTX in ovarian reserve and granulosa cells. Of note, as a major cause of DOR, the apoptosis of ovarian granulosa cells has been a consistent research focus concerning the etiology of DOR [ 33 ]. FOXO1 expression was found to be downregulated in granulosa cells from patients with poor ovarian reserve [ 34 ].…”

Effects of acupuncture treatment on microRNAs expression in ovarian tissues from Tripterygium glycoside-induced diminished ovarian reserve rats

“…MicroRNAs (miRNAs) are small non-coding RNAs, 20–25 nucleotides in length. Considering the critical role of miRNAs in regulating gene expression and interactive signaling events ( Flynt and Lai, 2008 ), their involvement in cumulus and granulosa cells has been demonstrated ( Chen et al, 2017 ; Hong et al, 2018 ; Kim et al, 2021a ; Wei et al, 2021 ). Despite the widespread application of miRNA sequencing ( Raza et al, 2022 ), the effect of acupuncture on miRNA profiling in DOR rats has not been fully elucidated.…”