Zhang Yang, Guo Hai, Ma Li, Chen Xiaoyong, Chen Guangdong
Department of Orthopedics, Shenzhen University General Hospital, Shenzhen 518055, People's Republic of China.
Department of Anesthesiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, 830000, People's Republic of China.
Cancer Manag Res. 2020 Sep 24;12:8975-8987. doi: 10.2147/CMAR.S269774. eCollection 2020.
This study was conducted to determine the expression and prognostic relevance of long intergenic non-protein coding RNA 839 (LINC00839) in osteosarcoma (OS) and to explore the detailed roles of LINC00839 in regulating OS cell activities and the mechanisms responsible for its cancer-promoting activity in OS.
The expression of LINC00839 in OS tissues and cell lines was determined by quantitative reverse transcription-polymerase chain reaction. After LINC00839 knockdown, cell counting kit-8 assay, flow cytometric analysis, transwell migration and invasion assay, and in vivo tumor xenograft assay were used to detect its effects on cellular processes in OS. Bioinformatics analyses were conducted to predict the putative miRNAs that target LINC00839. RNA immunoprecipitation assay, luciferase reporter assay, Western blotting analysis, and rescue assays were conducted to establish a relationship among LINC00839, microRNA-454-3p (miR-454-3p), and cellular mesenchymal to epithelial transition factor (c-Met) in OS.
LINC00839 was upregulated in OS tissues and cell lines. OS patients characterized with high LINC00839 expression exhibited shorter overall survival than patients with low LINC00839 expression. LINC00839 knockdown caused a significant reduction in OS cell proliferation, migration, and invasion in vitro. Furthermore, LINC00839 depletion inhibited OS tumor growth in vivo and induced apoptosis. Mechanistically, LINC00839 functions as a competitive endogenous RNA in OS by sponging miR-454-3p. c-Met was confirmed as a direct target gene for miR-454-3p in OS cells and was positively regulated by LINC00839 by competitively binding to miR-454-3p.
LINC00839 promoted the oncogenicity of OS by targeting the miR-454-3p/c-Met axis. The LINC00839/miR-454-3p/c-Met network may represent a potential target for OS therapy.
本研究旨在确定长链基因间非编码RNA 839（LINC00839）在骨肉瘤（OS）中的表达及预后相关性，并探讨LINC00839在调节OS细胞活性中的具体作用及其在OS中促癌活性的相关机制。
采用定量逆转录-聚合酶链反应检测LINC00839在OS组织和细胞系中的表达。LINC00839敲低后，使用细胞计数试剂盒-8检测、流式细胞术分析、Transwell迁移和侵袭检测以及体内肿瘤异种移植检测来检测其对OS细胞进程的影响。进行生物信息学分析以预测靶向LINC00839的假定微小RNA（miRNA）。通过RNA免疫沉淀检测、荧光素酶报告基因检测、蛋白质免疫印迹分析和拯救实验来建立OS中LINC00839、微小RNA-454-3p（miR-454-3p）和细胞间充质向上皮转化因子（c-Met）之间的关系。
LINC00839在OS组织和细胞系中上调。LINC00839高表达的OS患者总生存期短于LINC00839低表达的患者。LINC00839敲低导致OS细胞体外增殖、迁移和侵袭显著减少。此外，LINC00839缺失抑制体内OS肿瘤生长并诱导凋亡。机制上，LINC00839在OS中通过吸附miR-454-3p作为竞争性内源RNA发挥作用。c-Met被确认为OS细胞中miR-454-3p的直接靶基因，并且LINC00839通过竞争性结合miR-454-3p对其起正向调节作用。
LINC00839通过靶向miR-454-3p/c-Met轴促进OS的致癌性。LINC00839/miR-454-3p/c-Met网络可能代表OS治疗的潜在靶点。
