Guangdong Province Key Laboratory of Animal Nutritional Regulation, National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.
Guangdong Engineering & Research Center for Woody Fodder Plants, South China Agricultural University, Guangzhou, China.
Front Endocrinol (Lausanne). 2020 Mar 3;11:92. doi: 10.3389/fendo.2020.00092. eCollection 2020.
MicroRNAs (MiRNAs) play critical roles in the regulation of pituitary function. MiR-130a-3p has previously been found to be down-regulated in prolactinoma, but its roles in prolactin (PRL) regulation and the underlying mechanisms are still unclear. Heat stress has been shown to induce alteration of endocrine hormones and miRNAs expressions. However, there is limited information regarding the emerging roles of miRNAs in heat stress response. In this study, we transfected miR-130a-3p mimic into the pituitary adenoma cells (GH3 cells) to investigate the function of miR-130a-3p in regulating PRL. Our results showed that miR-130a-3p overexpression significantly decreased the PRL expression at both mRNA and protein levels. Subsequently, estrogen receptor α (ERα) was identified as a direct target of miR-130a-3p by bioinformatics prediction, luciferase reporter assay and western blotting assay. Furthermore, the inhibition of ERα caused by estrogen receptor antagonist significantly reduced the PRL expression. Overexpression of ERα rescued the suppressed expression of PRL caused by miR-130a-3p mimic. Besides, we also studied the effect of heat stress on PRL and miRNAs expressions. Interestingly, we found that heat stress reduced PRL and ERα expressions while it increased miR-130a-3p expression both and . Taken together, our results indicate that miR-130a-3p represses ERα by targeting its 3'UTR leading to a decrease in PRL expression, and miR-130a-3p is correlative with heat stress-induced PRL reduction, which provides a novel mechanism that miRNAs are involved in PRL regulation.
微小 RNA(miRNAs)在调节垂体功能中起着关键作用。先前发现 miR-130a-3p 在催乳素瘤中下调,但它在催乳素(PRL)调节中的作用及其潜在机制尚不清楚。热应激已被证明会诱导内分泌激素和 miRNAs 表达的改变。然而,关于 miRNAs 在热应激反应中的新兴作用的信息有限。在这项研究中,我们将 miR-130a-3p 模拟物转染到垂体腺瘤细胞(GH3 细胞)中,以研究 miR-130a-3p 在调节 PRL 中的功能。我们的结果表明,miR-130a-3p 的过表达显著降低了 PRL 在 mRNA 和蛋白质水平的表达。随后,通过生物信息学预测、荧光素酶报告基因测定和 Western blot 测定,确定雌激素受体 α(ERα)是 miR-130a-3p 的直接靶标。此外,雌激素受体拮抗剂对 ERα 的抑制显著降低了 PRL 的表达。ERα 的过表达挽救了 miR-130a-3p 模拟物引起的 PRL 表达抑制。此外,我们还研究了热应激对 PRL 和 miRNAs 表达的影响。有趣的是,我们发现热应激降低了 PRL 和 ERα 的表达,同时增加了 miR-130a-3p 的表达。综上所述,我们的结果表明,miR-130a-3p 通过靶向其 3'UTR 抑制 ERα,导致 PRL 表达降低,miR-130a-3p 与热应激诱导的 PRL 降低相关,这为 miRNAs 参与 PRL 调节提供了一种新的机制。