Lu Fei, Song Ying, Cui Shuo, Zhao Huafei, Chen Yuhua, Du Hao
Department of Microsurgery, The First Affiliated Hospital, College of Clinical Medicine of Henan University of Science and Technology Luoyang 471003, Henan Province, China.
Department of Pharmacy, The First Affiliated Hospital, College of Clinical Medicine of Henan University of Science and Technology Luoyang 471003, Henan Province, China.
Am J Transl Res. 2021 Nov 15;13(11):12588-12600. eCollection 2021.
The present study aimed to explore the biological functions and mechanism of long non-coding RNA myocardial infarction-associated transcript (MIAT) in melanoma progression. MIAT expression in melanoma tissue samples and cells was detected by quantitative real-time PCR. After gain-of-function and loss-of-function models were constructed, cell counting kit-8, EdU, and Transwell assays were employed to detect the proliferation, migration, and invasion of melanoma cells. catRAPID database was employed and RNA pull-down assay and RNA immunoprecipitation assay were utilized to verify, the binding relationship between MIAT and transcription factor 12 (TCF12). The binding of TCF12 to the promoter region of the gene of nuclear factor of activated T cells 5 (NFAT5) was verified by chromatin immunoprecipitation-quantitative PCR assay and dual-luciferase reporter gene assay. The regulatory effects of MIAT and TCF12 on NFAT5 expression were detected via Western blot. The results showed that MIAT expression was increased in melanoma tissues and cells, and was significantly associated with the AJCC stage and the differentiation of melanoma tissues. MIAT overexpression markedly facilitated melanoma cells' multiplication, migration, and invasion, while MIAT knockdown inhibited the multiplication, migration, and invasion. MIAT showed direct interaction with TCF12. MIAT promoted the binding of TCF12 to NFAT5 promoter region, thereby promoting NFAT5 transcription. In conclusion, MIAT promotes melanoma progression through recruiting TCF12 and its interaction with NFAT5.
本研究旨在探讨长链非编码RNA心肌梗死相关转录本（MIAT）在黑色素瘤进展中的生物学功能及机制。通过定量实时PCR检测黑色素瘤组织样本和细胞中MIAT的表达。构建功能获得和功能缺失模型后，采用细胞计数试剂盒-8、EdU和Transwell实验检测黑色素瘤细胞的增殖、迁移和侵袭能力。利用catRAPID数据库，并采用RNA下拉实验和RNA免疫沉淀实验验证MIAT与转录因子12（TCF12）之间的结合关系。通过染色质免疫沉淀-定量PCR实验和双荧光素酶报告基因实验验证TCF12与活化T细胞核因子5（NFAT5）基因启动子区域的结合。通过蛋白质免疫印迹法检测MIAT和TCF12对NFAT5表达的调控作用。结果显示，MIAT在黑色素瘤组织和细胞中的表达升高，且与AJCC分期及黑色素瘤组织的分化程度显著相关。MIAT过表达显著促进黑色素瘤细胞的增殖、迁移和侵袭，而MIAT敲低则抑制其增殖、迁移和侵袭。MIAT与TCF12存在直接相互作用。MIAT促进TCF12与NFAT5启动子区域的结合，从而促进NFAT5转录。综上所述，MIAT通过招募TCF12及其与NFAT5的相互作用促进黑色素瘤进展。
