LINC01116 靶向 miR-520a-3p 并影响 IL6R，通过 Jak-stat 信号通路促进骨肉瘤细胞的增殖和迁移。
LINC01116 targets miR-520a-3p and affects IL6R to promote the proliferation and migration of osteosarcoma cells through the Jak-stat signaling pathway.
机构信息
Department of Radiology, China-Japan Union Hospital of Jilin University, Changchun 130021, Jilin, China.
Department of Radiology, China-Japan Union Hospital of Jilin University, Changchun 130021, Jilin, China.
出版信息
Biomed Pharmacother. 2018 Nov;107:270-282. doi: 10.1016/j.biopha.2018.07.119. Epub 2018 Aug 8.
OBJECTIVE
The purpose of this study was to find out the important lncRNA-miRNA-mRNA axis and pathway in osteosarcoma (OS) through bioinformatics analysis and verify the biological functions of lncRNA/miRNA/mRNA in OS through in vitro and in vivo assays.
METHODS
The differential expression mRNAs and lncRNAs were identified through microarray analysis, and the altered pathways were identified by GSEA. The Pearson Coefficient was used to analyze the correlations between mRNAs and lncRNAs. Kaplan-Meier survival analysis was preformed using patient information in GEO database. Their target miRNAs were predicted by Targetscan and miRanda database and confirmed by dual luciferase reporter assay. QRT-PCR were utilized to detected the relative expressions of mRNAs, miRNAs and lncRNAs. The expressions of IL6R protein and pathway related proteins were detected by western blot. OS cell viability, migration and apoptosis were determined through MTT assay, Transwell assay and flow cytometry. Tumor formation in nude mice verified the influence of LINC01116 in vivo.
RESULTS
The Jak-stat signaling pathway was activated in OS tissues. LINC01116 expression was positively correlated with IL6R expression. MiR-520a-3p targeted the 3'-UTR of LINC01116 and IL6R. Lower expression levels of miR-520a-3p significantly correlated with shorter survival of patients. LINC01116 and IL6R were up-regulated while miR-520a-3p was down-regulated in OS. LINC01116 and IL6R promoted the viability and migration of OS cells, while miR-520a-3p acted as a tumor suppressor. MiR-520a-3p inhibitor could rescue the suppressive effects of si-LINC011116 and si-IL6R on OS development. The Jak-stat signaling pathway related proteins were also down-regulated by miR-520a-3p. Down-regulation of LINC01116 inhibited the tumor growth in nude mice.
CONCLUSION
LINC01116 up-regulated IL6R in OS through targeting miR-520a-3p, thus activating the Jak-stat signaling pathway and promoting the progression of OS.
目的
本研究通过生物信息学分析，找出骨肉瘤（OS）中重要的 lncRNA-miRNA-mRNA 轴和途径，并通过体外和体内实验验证 lncRNA/miRNA/mRNA 在 OS 中的生物学功能。
方法
通过微阵列分析鉴定差异表达的 mRNAs 和 lncRNAs，通过 GSEA 鉴定改变的途径。通过 Pearson 系数分析 mRNAs 和 lncRNAs 之间的相关性。使用 GEO 数据库中的患者信息进行 Kaplan-Meier 生存分析。使用 Targetscan 和 miRanda 数据库预测其靶 miRNAs，并通过双荧光素酶报告基因实验进行验证。使用 QRT-PCR 检测 mRNAs、miRNAs 和 lncRNAs 的相对表达水平。使用 Western blot 检测 IL6R 蛋白和通路相关蛋白的表达。通过 MTT 检测、Transwell 检测和流式细胞术测定 OS 细胞活力、迁移和凋亡。裸鼠肿瘤形成实验验证 LINC01116 在体内的影响。
结果
Jak-stat 信号通路在 OS 组织中被激活。LINC01116 的表达与 IL6R 的表达呈正相关。miR-520a-3p 靶向 LINC01116 和 IL6R 的 3'-UTR。miR-520a-3p 的低表达水平与患者的生存时间较短显著相关。LINC01116 和 IL6R 在 OS 中上调，而 miR-520a-3p 下调。LINC01116 和 IL6R 促进 OS 细胞的活力和迁移，而 miR-520a-3p 起肿瘤抑制作用。miR-520a-3p 抑制剂可挽救 si-LINC011116 和 si-IL6R 对 OS 发展的抑制作用。miR-520a-3p 还下调了 Jak-stat 信号通路相关蛋白。下调 LINC01116 抑制了裸鼠肿瘤的生长。
结论
LINC01116 通过靶向 miR-520a-3p 上调 OS 中的 IL6R，从而激活 Jak-stat 信号通路并促进 OS 的进展。
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