STAT3 activates the transcription of lncRNA NR2F1-AS1 to promote the progression of melanoma via regulating the miR-493-5p/GOLM1 axis
STAT3 activates the transcription of lncRNA NR2F1-AS1 to promote the progression of melanoma via regulating the miR-493-5p/GOLM1 axis
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DOI：
10.1002/jgm.3338
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发表时间：
2021-04-27
影响因子：
3.5
通讯作者：
Wang, Xiong
中科院分区：
文献类型：
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作者：
Bai, Mei;Wu, Zong-Zhou;Wang, Xiong
背景长非编码RNA(LncRNAs)在黑色素瘤的生物学过程中起着重要的调节作用。本研究旨在揭示NR2F1反义RNA 1(NR2F1-AS1)在黑色素瘤中的生物学功能及其可能的作用机制。方法采用实时定量聚合酶链式反应方法检测137例配对的原发黑色素瘤组织和相应的非肿瘤组织以及3个黑色素瘤细胞系中NR2F1-AS1和miR-493-5p的相对水平。统计分析NR2F1-AS1表达的临床意义。用JASPAR(http://jaspar.genereg.net).)鉴定了NR2F1-AS1启动子区域的STAT3结合基序用双荧光素酶报告和染色质免疫沉淀实验确定STAT3与NR2F1-AS1的相关性。用细胞计数试剂盒8(CCK-8)、EDU、Transwell和伤口愈合试验检测NR2F1-AS1对细胞增殖、迁移的影响。应用双荧光素酶报告和RNA下拉实验验证NR2F1-AS1、miR-493-5p和GOLM1之间的相互作用。结果NR2F1-AS1在黑色素瘤细胞中表达上调，miR-493-5p表达下调。STAT3诱导NR2F1-AS1过表达。NR2F1-AS1的高表达与黑色素瘤的分期和预后不良有关。应用CCK-8、EDU、Transwell和伤口愈合试验的功能研究表明，抑制NR2F1-AS1可减弱黑色素瘤细胞的增殖、迁移和侵袭能力。此外，NR2F1-AS1能够通过识别和结合miR-493-5p上调GOLM1的表达。此外，miR-493-5p的敲除明显逆转了NR2F1-AS1下调对黑色素瘤发生和发展的抑制作用。结论NR2F1-AS1通过靶向miR-493-5p/GOLM1轴在黑色素瘤的发生发展中发挥关键作用。
Background Long non-coding RNAs (lncRNAs) are vital regulators during the biological processes of melanoma. The present study aimed to uncover biological functions of lncRNA termed NR2F1 antisense RNA 1 (NR2F1-AS1) in melanoma and the potential mechanisms.Methods Relative levels of NR2F1-AS1 and miR-493-5p in a total of 137 paired primary melanoma tissues and corresponding non-tumor tissues, as well as three melanoma cell lines, were examined by a real-time polymerase chain reaction. The clinical significance of NR2F1-AS1 expression was analyzed statistically. The STAT3 binding motif in the promoter region of NR2F1-AS1 was identified by JASPAR (http://jaspar.genereg.net). The association between STAT3 and NR2F1-AS1 was determined by dual-luciferase reporter and chromatin immunoprecipitation assays. The effects of NR2F1-AS1 on cell proliferation, migration and were measured by cell counting kit-8 (CCK-8), Edu, transwell and wound healing assays. Dual-luciferase reporter and RNA pull-down assays were applied to validate the interaction among NR2F1-AS1, miR-493-5p and GOLM1. Furthermore, in vivo experiments were conducted to demonstrate the oncogenic role of NR2F1-AS1 in melanoma.Results Up-regulated NR2F1-AS1 and down-regulated miR-493-5p were detected in melanoma tumors and cells. The overexpression of NR2F1-AS1 was induced by STAT3. High NR2F1-AS1 expression was correlated to advanced tumor stage and poor prognosis of melanoma. Functional studies using CCK-8, Edu, transwell and wound healing assays revealed that the proliferative, migratory and invasive capacities of melanoma cells were attenuated by the by inhibition of NR2F1-AS1. Moreover, NR2F1-AS1 was able to up-regulate GOLM1 through recognizing and binding miR-493-5p. Furthermore, knockdown of miR-493-5p distinctly reversed these inhibitory effects of NR2F1-AS1 down-regulation on the tumorigenesis and progression of melanoma.Conclusions Our findings demonstrate a key role for NR2F1-AS1 in melanoma progression via targeting miR-493-5p/GOLM1 axis.