Choi Jiyun, Jang Eunjae, Jeong Haewon, Hwang Jinsu, Cho Hyong-Ho, Kim Byeong C, Jang Geupil, Jeong Han-Seong, Jang Sujeong
Department of Physiology, Chonnam National University Medical School, Hwasun-Gun, Jeollanamdo, 58128, Republic of Korea.
Jeonnam Bioindustry Foundation Biopharmaceutical Research Center, Hwasun-Gun, Jeollanamdo, 58141, Republic of Korea.
Mol Neurobiol. 2025 May 2. doi: 10.1007/s12035-025-05002-4.
MicroRNAs (miRNAs) are small non-coding RNAs that regulate various biological processes, including cell differentiation. Despite their potential, their role in promoting neuronal differentiation by targeting neuronal genes and modulating signaling pathways is poorly understood. In this study, we aimed to elucidate the functions of miR-937-3p-, miR-4536-3p-, and miR-4650-5p-inhibitors in the neuronal differentiation of SH-SY5Y cells. We also aimed to determine the underlying mechanisms via qPCR, luciferase assay, immunocytochemistry, and western blotting analysis. Our findings confirmed that miRNAs participated in neuronal differentiation and regulated the Wnt/MAPK signaling pathway. Specifically, we identified Netrin1 (NTN1), Drebrin1 (DBN1), and Netrin-G1 (NTNG1) as target genes of miR-937-3p, miR-4536-3p, and miR-4650-5p, respectively. The treatment with the miRNA inhibitors increased the expression levels of neuronal markers such as TUBB3, NEFH, NEFM, NEFL, and MAP2. It also enhanced the protein expression levels of Wnt and MAPK signaling. Therefore, the inhibitors of miR-937-3p, miR-4536-3p, and miR-4650-5p could promote neuronal differentiation by targeting neuronal genes and activating the Wnt/MAPK signaling pathway.
微小RNA（miRNA）是一类小的非编码RNA，可调节包括细胞分化在内的各种生物学过程。尽管它们具有潜在作用，但其通过靶向神经元基因和调节信号通路来促进神经元分化的作用仍知之甚少。在本研究中，我们旨在阐明miR-937-3p、miR-4536-3p和miR-4650-5p抑制剂在SH-SY5Y细胞神经元分化中的功能。我们还旨在通过定量聚合酶链反应（qPCR）、荧光素酶测定、免疫细胞化学和蛋白质免疫印迹分析来确定其潜在机制。我们的研究结果证实，miRNA参与神经元分化并调节Wnt/丝裂原活化蛋白激酶（MAPK）信号通路。具体而言，我们分别确定了神经细胞粘附分子1（Netrin1，NTN1）、肌动蛋白结合蛋白1（Drebrin1，DBN1）和神经细胞粘附分子G1（Netrin-G1，NTNG1）为miR-937-3p、miR-4536-3p和miR-4650-5p的靶基因。用miRNA抑制剂处理可增加神经元标志物如微管蛋白β-3（TUBB3）、神经丝重链（NEFH）、神经丝中链（NEFM）、神经丝轻链（NEFL）和微管相关蛋白2（MAP2）的表达水平。它还增强了Wnt和MAPK信号的蛋白质表达水平。因此，miR-937-3p、miR-4536-3p和miR-4650-5p抑制剂可通过靶向神经元基因并激活Wnt/MAPK信号通路来促进神经元分化。
