Department of Gynecology and Obstetrics, Reproductive Medical Center, Shanghai Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Department of Dermatology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Aging (Albany NY). 2020 Nov 16;12(22):22759-22775. doi: 10.18632/aging.103909.
Melanoma is a skin malignancy with a high mutation frequency of genetic alterations. MicroRNA (miR)-200b-3p is involved in various cancers, while in melanoma its bio-function remains unknown. In this study, we found that miR-200b-3p was down-regulated in melanoma tissues and cell lines compared to benign nevus cells. Overexpression of miR-200b-3p significantly inhibited the proliferation and invasion of melanoma cells. According to bioinformatics analysis and sequencing data, we supposed that SMAD family member 2 (SMAD2) was the target gene and nuclear enriched abundant transcript 1 (NEAT1) was the upstream long non-coding RNA (lncRNA) of miR-200b-3p. These predictions were verified by western blotting and quantitative real-time reverse transcription PCR (RT-qPCR). Luciferase reporter assays revealed that NEAT1 up-regulated SMAD2 by directly sponging miR-200b-3p. and , we demonstrated that both NEAT1 and SMAD2 could promote the proliferation and invasion of melanoma cells, and these effects were reversed by up-regulating miR-200b-3p. In addition, NEAT1/miR-200b-3p/SMAD2 axis promoted melanoma progression by activating EMT signaling pathway and immune responses. Taken together, the NEAT1/miR-200b-3p/SMAD2 signaling pathway promotes melanoma via activation of EMT, cell invasion and is related with immune responses, which provides new insights into the molecular mechanisms and therapeutic targets for melanoma.
黑色素瘤是一种具有高基因突变频率的皮肤恶性肿瘤。MicroRNA (miR)-200b-3p 参与多种癌症，而在黑色素瘤中其生物功能尚不清楚。在本研究中，我们发现 miR-200b-3p 在黑色素瘤组织和细胞系中较良性痣细胞下调。miR-200b-3p 的过表达显著抑制黑色素瘤细胞的增殖和侵袭。根据生物信息学分析和测序数据，我们推测 SMAD 家族成员 2 (SMAD2) 是 miR-200b-3p 的靶基因，核富集丰富转录物 1 (NEAT1) 是 miR-200b-3p 的上游长非编码 RNA (lncRNA)。这些预测通过 Western blot 和定量实时逆转录 PCR (RT-qPCR) 得到验证。荧光素酶报告基因实验表明，NEAT1 通过直接海绵吸附 miR-200b-3p 上调 SMAD2。此外，我们表明 NEAT1 和 SMAD2 均可促进黑色素瘤细胞的增殖和侵袭，而通过上调 miR-200b-3p 可逆转这些作用。此外，NEAT1/miR-200b-3p/SMAD2 轴通过激活 EMT 信号通路和免疫反应促进黑色素瘤的进展。总之，NEAT1/miR-200b-3p/SMAD2 信号通路通过激活 EMT、细胞侵袭促进黑色素瘤的进展，并与免疫反应有关，为黑色素瘤的分子机制和治疗靶点提供了新的见解。
