Aug 2026· International Journal of Biological Macromolecules· Vol 380, pp.
154055
· 0 citations· 69 references
Medicine
TL;DR
A bta-miR-380-3p-SOAT1 regulatory axis linking post-transcriptional control with cholesterol metabolism, preadipocyte expansion, and IMF deposition is revealed, providing mechanistic insight and a potential molecular target for improving marbling and meat quality traits in cattle breeding programs.
Abstract
Intramuscular fat (IMF) deposition significantly impacts beef quality, yet microRNA (miRNA)-mediated mechanisms regulating bovine intramuscular preadipocyte development are not fully understood. This study aimed to elucidate the role of bta-miR-380-3p in preadipocyte proliferation, apoptosis, and adipogenic differentiation, identifying sterol O-acyltransferase 1 (SOAT1) as its functional target. Bta-miR-380-3p exhibited dynamic expression during adipogenesis and was highly expressed in adult bovine IMF. Gain- and loss-of-function assays showed that bta-miR-380-3p promoted DNA synthesis, cell viability, and cell-cycle progression, and reduced apoptosis-related signaling. Consistently, western blot analysis confirmed increased PCNA and CDK1 and reduced P21 after bta-miR-380-3p overexpression, whereas inhibition produced the opposite pattern. Bta-miR-380-3p also enhanced lipid droplet accumulation and upregulated adipogenic markers, with western blotting confirming increased PPARγ, FABP4, and SREBP1 protein abundance. Bioinformatics prediction, expression validation, and dual-luciferase reporter assays demonstrated that bta-miR-380-3p directly targets the SOAT1 3'UTR and negatively regulates SOAT1 expression. Functional manipulation of SOAT1 further revealed its involvement in triglyceride accumulation, adipogenic marker regulation, and proliferation-associated remodeling. Collectively, these findings reveal a bta-miR-380-3p-SOAT1 regulatory axis linking post-transcriptional control with cholesterol metabolism, preadipocyte expansion, and IMF deposition. These results provide mechanistic insight and a potential molecular target for improving marbling and meat quality traits in cattle breeding programs.
The findings indicate that the Bta-miR-146a/SMAD4/TGFβ/MAPK axis is a key regulator of SC proliferation and apoptosis, offering insights into the molecular mechanisms underlying bovine spermatogenesis and potential targets for improving reproductive performance.
Qi-Wen Lu, Quan-Heng Guo, Yanlong Zhou et al.· International Journal of Mol...· 0 citations
MicroRNAs (miRNAs) play critical roles in regulating follicular development and atresia by modulating autophagy, apoptosis, and steroidogenesis in granulosa cells (GCs). However, the role of specific miRNAs in these processes remains poorly understood. In this study, we focused on miR-6615-3p to explore its function and molecular mechanism in chicken Prehierarchical follicular granulosa cells (PFGCs). The expression of miR-6615-3p in chicken PFGCs was confirmed by qRT-PCR. Dual-luciferase reporter assays identified the androgen receptor (AR) gene as a direct target of miR-6615-3p. Functional analyses revealed that miR-6615-3p suppressed AR expression, promoted apoptosis, and inhibited both autophagy and steroidogenesis in PFGCs. Together, these results suggest that miR-6615-3p downregulates AR and is associated with reduced MDM2 expression, enhanced p53 activation, and decreased BCL2 expression. This indicates that it may regulate cell fate and steroid hormone synthesis, at least in part, through the MDM2-p53-BCL2 signaling pathway. This study provides new insights into the molecular mechanisms underlying chicken follicular development and highlights miR-6615-3p as a potential molecular marker for improving reproductive performance in poultry.
Background Diminished ovarian reserve (DOR) is a common cause of female infertility. Prior high-throughput sequencing showed specific follicular fluid exosomal miRNA profiles in DOR patients, implicating exosomal miRNAs in DOR pathogenesis. Using miRNA PCR arrays, we found miR-483-3p was significantly upregulated in DOR follicular fluid exosomes. Bioinformatics and experiments indicated METTL3 as a potential miR-483-3p target, suggesting excess miR-483-3p exerts cytotoxicity and suppresses METTL3-mediated N6-methyladenosine (m6A) modification. This study thus investigated miR-483-3p effects on METTL3 expression, m6A levels, cell proliferation, and mitochondrial function in ovarian granulosa cells (GCs). Methods The targeting interaction between miR-483-3p and METTL3 was confirmed using a dual-luciferase reporter gene assay. qRT-PCR, immunofluorescence, Western blot, and Dot blot were used to evaluate the effects of miR-483-3p mimics, cyclophosphamide (CTX), and STM2457 on METTL3 protein expression and m6A modification levels in KGN cells. Additionally, Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2’-deoxyuridine (EdU) assays, along with mitochondrial membrane potential (MMP) measurements, were used to examine the impact of these treatments on KGN cell proliferation and mitochondrial function. Results miR-483-3p directly targeted the 3′ untranslated region (3′UTR) of METTL3, and its mimics significantly suppressed METTL3 gene expression. In contrast, CTX robustly promoted METTL3 gene expression, accompanied by a substantial elevation in global m6A levels. STM2457 treatment also showed a similar trend. But miR-483-3p overexpression modestly inhibited METTL3 protein and m6A levels. Regarding cellular phenotypes, miR-483-3p overexpression, CTX, and STM2457 all exerted significant inhibitory effects on KGN cells. All three treatments consistently suppressed the cell viability of KGN cells, reduced the EdU-positive cell ratio, and decreased MMP levels. Conclusion This study provides preliminary evidence suggesting a specific regulatory interaction between miR-483-3p and METTL3. We demonstrate that aberrantly high expression of miR-483-3p significantly suppresses METTL3 mRNA levels but only moderately reduces METTL3 protein and m6A levels, suggesting the involvement of complex post-transcriptional regulatory mechanisms. Furthermore, overaccumulation of miR-483-3p markedly inhibits the proliferation of ovarian GCs and impairs their mitochondrial function. Further investigation into the role of the candidate miR-483-3p-METTL3-m6A axis in DOR will be conducted using in vivo models alongside an expanded cohort of clinical samples.
Findings reveal a novel epigenetic regulatory axis (novel-miR-357/S100A2 mRNA) involved in early muscle development and provide an important molecular mechanism for deeper understanding of the epigenetic regulatory processes underlying heterosis in cattle.
Gaoqing Xu, Chunqi Hou, He Ding et al.· Animal Genetics· 0 citations
Long non-coding RNA LINC01419 has been implicated in ferroptosis in various pathological conditions, yet its role in type 2 diabetes mellitus (T2DM) associated vascular endothelial dysfunction remains unclear. This study investigated the mechanism by which LINC01419 modulates ferroptosis in human vascular endothelial cells under high-glucose conditions. A high-glucose cellular model was established to evaluate cell proliferation, ferroptosis, and oxidative stress. Bioinformatics predictions and dual-luciferase reporter assays confirmed direct interactions between LINC01419 and miR-132-3p, and between miR-132-3p and ubiquitin-specific peptidase 38 (USP38). Quantitative real-time PCR validated regulatory interactions within the LINC01419–miR-132-3p–USP38 axis. Cell viability, reactive oxygen species levels, and ferroptosis markers were assessed via CCK-8, ROS detection, and Western blotting. Results showed that high glucose upregulated LINC01419 and USP38, downregulated miR-132-3p, elevated ROS, suppressed proliferation, and induced ferroptosis, as evidenced by increased ACSL4 and COX2 and decreased FTH1 and GPX4. LINC01419 acted as a competing endogenous RNA for miR-132-3p, and both molecules oppositely regulated USP38 expression. Notably, USP38 knockdown reversed high-glucose-induced abnormalities, restoring proliferation and ferroptosis-related protein expression. These findings reveal that the LINC01419–miR-132-3p–USP38 axis mediates ferroptosis in vascular endothelial cells under hyperglycemia, and suggest USP38 as a potential therapeutic target for T2DM.
Ke-Ke Tang, Wen-Ji Gao, Xin-Xing Wo et al.· Scientific Reports· 0 citations
MicroRNAs are important post-transcriptional regulators of intracellular signaling pathways that govern cell proliferation, apoptosis, and cell-cycle progression. However, whether miR-424-3p regulates PIP5K1C-associated AKT signaling in granulosa cells remains unclear. In this study, we investigated the molecular mechanism of miR-424-3p in goat granulosa cells (GCs). RT-qPCR analysis showed that miR-424-3p was more highly expressed in large follicles than in small follicles. Gain- and loss-of-function assays revealed that miR-424-3p inhibited GC proliferation, reduced the proportion of cells in S phase, and promoted apoptosis. Western blot analysis showed that miR-424-3p decreased the protein levels of CDK6 and CCND1. Dual-luciferase reporter assays and Western blotting identified BCL2 and PIP5K1C as direct targets of miR-424-3p. Functional rescue experiments further demonstrated that PIP5K1C overexpression promoted GC proliferation, increased CCND1 expression, and partially counteracted the inhibitory effects of miR-424-3p. Mechanistically, miR-424-3p suppressed AKT phosphorylation, whereas PIP5K1C enhanced AKT phosphorylation, suggesting that the miR-424-3p/PIP5K1C axis regulates GC fate through AKT signaling. These findings identify a previously unrecognized miR-424-3p/PIP5K1C/AKT regulatory axis that links post-transcriptional gene regulation to AKT-dependent granulosa cell proliferation and apoptosis.
