College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.
Institute of Reproduction and Metabolism, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.
In Vitro Cell Dev Biol Anim. 2021 Sep;57(8):817-824. doi: 10.1007/s11626-021-00611-4. Epub 2021 Oct 28.
Both microRNA-7a (miR-7a) and LIM-homeodomain transcription factor ISL1 are important factors regulating insulin transcription and secretion, but the functional relationship and the interacting mechanisms between miR-7a and ISL1 in pancreatic islet β-cells remain unknown. The aims of this study were thus to identify the potential interactions and signaling communication between miR-7a and ISL1 in regulating insulin transcription and secretion in the cultured NIT-1 cells. The results show that miR-7a inhibitor upregulates Isl-1 and insulin gene expressions, and the insulin secretion. Whereas miR-7a mimics inhibit ISL1 and insulin gene expressions, and decreases the insulin secretion. Furthermore, we identified the target gene of miR-7a using dual-luciferase reporter assay, and the results demonstrate that Raf1 and Mapkap1 is a direct target gene of miR-7a, modeling RAF1/MEK/ERK1/2 and mTORC2/AKT signaling pathway to regulate Isl1 expression, and thus influencing insulin expression and secretion. Our results indicate that therapeutic inhibition of miR-7a function could be of relevance for preserving the function of pancreatic β-cells during the course of diabetes development, implicating miR-7, ISL1, and/or the connecting molecules may act as novel targets for pharmacological or gene therapy in diabetes and related metabolic disease, although much detailed studies are required in the further study.
miR-7a 和 LIM 同源结构域转录因子 ISL1 都是调节胰岛素转录和分泌的重要因素,但 miR-7a 和 ISL1 在胰岛β细胞中的功能关系和相互作用机制尚不清楚。因此,本研究旨在鉴定 miR-7a 和 ISL1 在调节培养的 NIT-1 细胞中胰岛素转录和分泌中的潜在相互作用和信号通讯。结果表明,miR-7a 抑制剂上调 Isl-1 和胰岛素基因的表达,并增加胰岛素的分泌。而 miR-7a 模拟物则抑制 ISL1 和胰岛素基因的表达,并减少胰岛素的分泌。此外,我们通过双荧光素酶报告基因检测鉴定了 miR-7a 的靶基因,结果表明 Raf1 和 Mapkap1 是 miR-7a 的直接靶基因,通过调节 RAF1/MEK/ERK1/2 和 mTORC2/AKT 信号通路来调节 Isl1 表达,从而影响胰岛素的表达和分泌。我们的结果表明,抑制 miR-7a 的功能可能与在糖尿病发展过程中保护胰岛β细胞的功能有关,这暗示着 miR-7、ISL1 及其连接分子可能成为糖尿病及相关代谢性疾病的药理学或基因治疗的新靶点,尽管还需要进一步的详细研究。