背景:研究长链非编码RNA(lncRNA)在骨肉瘤进展中的潜在作用。
方法:用RNA-seq筛选候选lncRNAs并用定量实时PCR确认。使用MTS,transwell分析,和流式细胞仪分析,测定过表达的lnc-SELPLG-2:1对细胞功能的影响。免疫组织化学染色,荧光原位杂交,和荧光素酶报告基因测定用于使用肿瘤模型在体内和体外评估lnc-SELPLG-2:1的潜在机制。此外,使用功能细胞分析确定hsa-miR-10a-5p过表达对SaOS2细胞功能的影响。使用反应测试来确认lnc-SELPLG-2:1海绵hsa-miR-10a-5p促进BTRC表达以调节骨肉瘤的机制。
结果:与正常细胞和骨髓样本相比,Lnc-SELPLG-2:1在骨肉瘤中高表达。抑制lnc-SELPLG-2:1加速细胞凋亡和抑制细胞增殖,迁移,和入侵,而lnc-SELPLG-2:1过表达具有相反的作用。此外,在体内模型中抑制lnc-SELPLG-2:1可减小肿瘤大小并抑制细胞迁移相关蛋白的表达。预测,双荧光素酶测定,应答检测结果显示hsa-miR-10-5p和BTRC参与lnc-SELPLG-2:1级联反应。与lnc-SELPLG-2:1不同,hsa-hsa-miR-10a-5p具有相反的表达和功能。lnc-SELPLG-2:1与hsa-hsa-miR-10a-5p的竞争性结合阻止了BTRC的miRNA介导的降解,从而激活VIM的表达式,MMP9和MMP2促进骨肉瘤细胞增殖,迁移,和入侵,抑制细胞凋亡。
结论:Lnc-SELPLG-2:1是骨肉瘤的致癌激活剂,其功能通过hsa-miR-10a-5p/BTRC级联进行。
BACKGROUND: To investigate the potential role of Long Non-coding RNAs (lncRNAs) in the progression of osteosarcoma.
METHODS: The candidate lncRNAs were screened with RNA-seq and confirmed with quantitative real-time PCR. Using MTS, transwell assay, and flow cytometric analysis, the effects of overexpressed lnc-SELPLG-2:1 on cell functions were determined. Immunohistochemical staining, fluorescence in situ hybridization, and luciferase reporter assay were used to evaluate the potential mechanism of lnc-SELPLG-2:1 in vivo and in vitro using a tumor model. Moreover, the effects of overexpression of hsa-miR-10a-5p on the functions of SaOS2 cells were determined using functional cell analysis. A response test was used to confirm the mechanism by which lnc-SELPLG-2:1 sponge hsa-miR-10a-5p promotes the expression of
BTRC to regulate osteosarcoma.
RESULTS: Lnc-SELPLG-2:1 was highly expressed in osteosarcoma compared to normal cells and bone and marrow samples. Inhibition of lnc-SELPLG-2:1 accelerated cell apoptosis and suppressed cell proliferation, migration, and invasion, whereas lnc-SELPLG-2:1 overexpression had the opposite effect. Moreover, inhibiting lnc-SELPLG-2:1 in an in vivo model decreased tumor size and suppressed the expression of cell migration-related proteins. The prediction, dual luciferase assay, and response test results indicated that hsa-miR-10-5p and
BTRC were involved in the lnc-SELPLG-2:1 cascade. Unlike lnc-SELPLG-2:1, hsa-hsa-miR-10a-5p had opposite expression and function. Competitive binding of lnc-SELPLG-2:1 to hsa-hsa-miR-10a-5p prevented
BTRC from miRNA-mediated degradation, thereby activating the expression of VIM, MMP9, and MMP2, promoting osteosarcoma cell proliferation, migration, and invasion, and inhibiting apoptosis.
CONCLUSIONS: Lnc-SELPLG-2:1 is an oncogenesis activator in osteosarcoma, and its functions are performed via hsa-miR-10a-5p /
BTRC cascade.