Department of Dermatology, School of Medicine, Guangzhou First People's Hospital, South China University of Technology, Guangzhou, Guangdong 510180, P.R. China.
Mol Med Rep. 2021 Jan;23(1). doi: 10.3892/mmr.2020.11717. Epub 2020 Nov 25.
Micro (mi)RNAs serve crucial roles in cancer development although little is known about their cellular mechanisms in the pathogenesis of melanoma. The present study explored the regulatory roles of miR‑18a‑5p in melanoma cell proliferation, apoptosis and autophagy, in addition to its target gene in melanoma cells. miRNA and ephrin receptor A7 (EPHA7) mRNA were analyzed by reverse transcription‑quantitative PCR. Cell Counting Kit‑8 and colony formation assays were performed to examine the cell proliferation rate. Hoechst staining and flow cytometry were performed to investigate cell apoptosis. Western blotting was used to estimate the abundance of proteins. Dual-luciferase reporter assay verified the binding of miRNA with target gene sequences. Melanoma tissues and cell lines exhibited markedly elevated miR‑18a‑5p expression. miR‑18a‑5p inhibitor inhibited proliferation rates, and triggered apoptosis and autophagy marker protein expression in WM266‑4 and A375 cells. It also negatively regulated EPHA7 expression in WM266‑4 and A375 cells by directly binding at the 3'‑untranslated region of EPHA7. miR‑18a‑5p mimics reversed the EPHA7 overexpression‑induced suppression of proliferation, and the EPHA7 overexpression‑induced promotion of apoptosis and autophagy. miR‑18a‑5p triggered proliferation of melanoma cells and inhibited apoptosis and autophagy by directly targeting and inhibiting EPHA7 expression. Thus, the present study aided our understanding of miRNA‑mediated melanoma pathogenesis.
微小 RNA(miRNAs)在癌症发展中发挥着至关重要的作用,尽管人们对其在黑色素瘤发病机制中的细胞机制知之甚少。本研究探讨了 miR-18a-5p 在黑色素瘤细胞增殖、凋亡和自噬中的调节作用,以及其在黑色素瘤细胞中的靶基因。采用逆转录定量 PCR 分析 miRNA 和 Ephrin 受体 A7(EPHA7)mRNA。通过细胞计数试剂盒-8 和集落形成实验检测细胞增殖率。采用 Hoechst 染色和流式细胞术检测细胞凋亡。采用 Western blot 法估计蛋白丰度。双荧光素酶报告基因实验验证 miRNA 与靶基因序列的结合。黑色素瘤组织和细胞系表现出明显升高的 miR-18a-5p 表达。miR-18a-5p 抑制剂抑制 WM266-4 和 A375 细胞的增殖率,并触发细胞凋亡和自噬标志物蛋白表达。它还通过直接结合 EPHA7 的 3'-非翻译区负调控 WM266-4 和 A375 细胞中的 EPHA7 表达。miR-18a-5p 模拟物逆转了 EPHA7 过表达诱导的增殖抑制,以及 EPHA7 过表达诱导的凋亡和自噬促进。miR-18a-5p 通过直接靶向和抑制 EPHA7 表达,触发黑色素瘤细胞的增殖并抑制凋亡和自噬。因此,本研究有助于我们理解 miRNA 介导的黑色素瘤发病机制。