Xinxiang Central Hospital, Xinxiang 453000, Henan, China.
Aging (Albany NY). 2021 Feb 1;13(4):5297-5311. doi: 10.18632/aging.202450.
In this study, we investigated the role of lncRNA MIR205HG in melanomagenesis. Quantitative real-time PCR (qRT-PCR) analysis showed that MIR205HG levels were significantly upregulated in melanoma cell lines compared to normal human melanocytes. Similarly, MIR205HG levels were significantly higher melanoma tissues than adjacent normal skin tissues (n=30). CCK-8 and flow cytometry assays showed that MIR205HG knockdown significantly decreased the viability of melanoma cells. Dual luciferase reporter and RNA pull-down assays confirmed that MIR205HG directly binds to microRNA (miR)-299-3p. Targetscan analysis and dual luciferase reporter assays showed that miR-299-3p directly binds to the 3'UTR of VEGFA mRNA. Wound healing and transwell invasion assays showed that MIR205HG knockdown decreased migration and invasiveness of melanoma cells, and these effects were reversed by treatment with miR-299-3p inhibitor. MIR205HG-silenced melanoma cells showed increased miR-299-3p expression and lower levels of both VEGFA mRNA and protein. Tumor volumes were significantly smaller in nude mice xenografted with MIR205HG knockdown melanoma cells than the controls. These results demonstrate that MIR205HG supports melanoma growth via the miR-299-3p/VEGFA axis. This makes MIR205HG a potential therapeutic target for the treatment of melanoma.
在这项研究中，我们调查了长非编码 RNA MIR205HG 在黑色素瘤发生中的作用。实时定量 PCR (qRT-PCR) 分析显示，与正常人黑色素细胞相比，MIR205HG 在黑色素瘤细胞系中的水平显著上调。同样，与相邻正常皮肤组织相比，MIR205HG 在黑色素瘤组织中的水平显著升高(n=30)。CCK-8 和流式细胞术检测表明，MIR205HG 敲低显著降低了黑色素瘤细胞的活力。双荧光素酶报告和 RNA 下拉实验证实，MIR205HG 可直接结合 microRNA (miR)-299-3p。Targetscan 分析和双荧光素酶报告实验表明，miR-299-3p 可直接结合 VEGFA mRNA 的 3'UTR。划痕愈合和 Transwell 侵袭实验表明，MIR205HG 敲低可降低黑色素瘤细胞的迁移和侵袭能力，而 miR-299-3p 抑制剂处理可逆转这些作用。沉默 MIR205HG 的黑色素瘤细胞显示出 miR-299-3p 表达增加，VEGFA mRNA 和蛋白水平降低。与对照组相比，用沉默 MIR205HG 的黑色素瘤细胞异种移植的裸鼠肿瘤体积明显更小。这些结果表明，MIR205HG 通过 miR-299-3p/VEGFA 轴支持黑色素瘤的生长。这使得 MIR205HG 成为治疗黑色素瘤的潜在治疗靶点。
