miR-192对猪子宫内膜上皮细胞生物学功能的调控作用

Regulatory effects of miR-192 on porcine endometrial epithelial cells

摘要

目的 miRNAs是一种内源性非编码RNA,广泛参与多种生理过程的转录后基因调控,包括胚胎附植和发育过程。目前关于miR-192的研究主要集中于癌症方面,在动物繁殖上的研究鲜有报道。基于课题组前期研究的基础,推测miR-192可能是猪胚胎附植的潜在调控因子。方法 本研究使用miR-192 mimics、mimics-NC、inhibitor和inhibitor -NC转染猪子宫内膜上皮细胞(pEECs),通过伤口愈合试验、qRT-PCR和Western blot检测miR-192对pEECs增殖、凋亡、迁移及相关标志基因和胚胎附植相关因子的影响。结果 过表达miR-192促进抑癌因子cyclin-dependent kinase inhibitor 1A(P21)和tumor protein p53(P53)基因mRNA的表达并抑制pEECs的增殖;过表达miR-192促进凋亡基因cysteine-dependent aspartate-directed protease-3(Caspase-3)基因mRNA和蛋白的表达;过表达miR-192促进matrix metalloproteinase 2(MMP2)和matrix metalloproteinase 9(MMP9)基因mRNA的表达和细胞迁移;过表达miR-192不同程度地促进胚胎附植相关因子transforming growth factor-beta(TGF-β)、leukemia inhibitory factor(LIF)、epidermal growth factor receptor(EGFR)和integrin beta 3(ITGB3)基因mRNA的表达;抑制miR-192表达结果则相反。结论 miR-192抑制pEECs的增殖,促进其凋亡、迁移过程并有利于胚胎附植相关因子TGF-β、LIF、EGFR和ITGB3的表达。

Abstract

Objective MicroRNAs (miRNAs) are endogenous non-coding RNAs that are widely involved in post-transcriptional gene regulation in various physiological processes, including embryo implantation and development. Studies on miR-192 have primarily focused on its role in cancer, whereas few studies have focused on its relevance in animal reproduction. Based on the previous research findings of the research group, it is hypothesized that miR-192 may be a potential regulatory factor in porcine embryo implantation. Methods In this study, porcine endometrial epithelial cells (pEECs) were transfected with miR-192 mimics, mimic NCs, inhibitors, and inhibitor NCs, and investigated the effects of miR-192 on the proliferation, apoptosis, migration, and embryonic attachment-related factors of pEECs in a wound-healing assay, qRT-PCR, and western blot analysis. Results miR-192 increased the mRNA expression of the oncogenes cyclin-dependent kinase inhibitor 1A(P21) and tumor protein p53(P53) while inhibiting the proliferation of pEECs. It also promoted the mRNA and protein expression levels of cysteine-dependent aspartate-directed protease-3(caspase-3) and the mRNA expression of matrix metalloproteinase 2(MMP2) and matrix metalloproteinase 9(MMP9) while enhancing cell migration. miR-192 promoted the mRNA expression of the embryonic attachment-related factors transforming growth factor beta (TGF-β), leukemia inhibitory factor (LIF), epidermal growth factor receptor (EGFR), and integrin beta 3 (ITGB3). Meanwhile, the inhibition of miR-192 yielded contrasting results. Conclusion miR-192 inhibited the proliferation and promoted the apoptosis and migration of pEECs. It also promoted the expression of TGF-β, LIF, EGFR, and ITGB3, which are associated with embryo implantation.

关键词

miR-192 / 猪 / 子宫内膜上皮细胞 / 胚胎附植

Key words

miR-192 / pig / endometrial epithelial cells / embryonic implantation

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基金资助

新疆生产建设兵团重点研发计划(2022AB012,2024AB015),新疆维吾尔自治区科技计划项目(2022LQ01003,2022TSYCCX0047),新疆生产建设兵团科技计划项目(2025YD015)