MiR-214-3p promotes proliferation and inhibits estradiol synthesis in porcine granulosa cells

Abstract

Background Granulosa cells (GCs) proliferation and estradiol synthesis significantly affect follicular development. The miR-214-3p expression in the ovarian tissues of high-yielding sows is higher than that in low-yielding sows, indicating that miR-214-3p may be involved in sow fertility. However, the functions and mechanisms of miR-214-3p on GCs are unclear. This study focuses on miR-214-3p in terms of the effects on GCs proliferation and estradiol synthesis. Results Our findings revealed that miR-214-3p promotes proliferation and inhibits estradiol synthesis in porcine GCs. MiR-214-3p can increase the percentage of S-phase cells, the number of EdU labeled positive cells, and cell viability. However, E2 concentration was reduced after miR-214-3p agomir treatment. We also found that miR-214-3p up-regulates the expression of cell cycle genes including cell cycle protein B (Cyclin B), cell cycle protein D (Cyclin D), cell cycle protein E (Cyclin E), and cyclin-dependent kinase 4 (CDK4) at the transcription and translation levels, but down-regulates the mRNA and protein levels of cytochrome P450 family 11 subfamily A member 1 (CYP11A1), cytochrome P450 family 19 subfamily A member 1 (CYP19A1), and steroidogenic acute regulatory protein (StAR) (i.e., the key enzymes in estradiol synthesis). On-line prediction, bioinformatics analysis, a luciferase reporter assay, RT-qPCR, and Western blot results showed that the target genes of miR-214-3p in proliferation and estradiol synthesis are Mfn2 and NR5A1, respectively. Conclusions Our findings suggest that miR-214-3p plays an important role in the functional regulation of porcine GCs and therefore may be a target gene for regulating follicular development.

Circadian clock gene BMAL1 controls testosterone production by regulating steroidogenesis‐related gene transcription in goat Leydig cells

“…Lysates from goat testes and LCs and western blot analysis procedures were performed as previously described (Gao et al, 2019; Shi et al, 2020). Goat testes and LCs were harvested at the indicated time points.…”

Lnc2300 is a cis‐acting long noncoding RNA of CYP11A1 in ovarian granulosa cells

“…However, only two miRNAs (miR‐214 and miR‐101) have been shown to influence the expression of CYP11A1 in large livestock without deeply mechanism investigation (X. An et al, 2020; Shi et al, 2020). In this study, we found that miR‐365‐3p induced GC apoptosis by directly targeting and inhibiting the expression of CYP11A1, indicating that CYP11A1 is a novel direct functional target of miR‐365‐3p.…”

Effects of Curcumin on Mitochondrial Function, Endoplasmic Reticulum Stress, and Mitochondria-Associated Endoplasmic Reticulum Membranes in the Jejunum of Oxidative Stress Piglets

“…Real-Time PCR (RT-qPCR) Experiment. Relative mRNA levels of MAM-related genes (mitofusin 1, Mfn1 and mitofusin 2, Mfn2) were assessed according to the work of Shi et al 26 Total RNA extraction was performed through an RNA extraction kit (Invitrogen Bioscience), and cDNA preparation and RT-qPCR were performed using a cDNA synthesis kit and ChamQ SYBR qPCR Master Mix (Vazyme, Nanjing, China). Supporting Table S2 enlists the sequence of primers.…”