Abstract:
Background: Long non-coding RNAs (lncRNAs) are associated with multiple head and neck tumors and play important roles in cancer. This study explored the molecular mechanism of Linc00662 in hypopharyngeal squamous cell carcinoma (HSCC). Methods: Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect gene expression in HSCC tissues. The viability and proliferation of tumor cells were measured using CCK-8 assays. HSCC cell apoptosis was measured using flow cytometry and western blotting. Cell stemness was examined using the sphere formation assay. A xenograft tumor model was established to investigate the role of Linc00662 in vivo. Results: The expression level of Linc00662 in HSCC tissues was significantly higher than that in adjacent normal tissues. The expression of Linc00662 had no significant relationship with the tumor stage. Patients with high Linc00662 expression were found to have shorter overall survival than those with low Linc00662 expression. Linc00662 over-expression promoted cell viability and inhibited apoptosis. Using online databases and a dual luciferase reporter, miR-15b-5p was confirmed as a potential downstream sponge of Linc00662. Moreover, Linc00662 was negatively associated with miR-15b-5p in HSCC cells. Depletion of miR-15b-5p can reverse the function of Linc00662 in vivo and in vitro. Furthermore, Linc00662 promotes tumor growth, which was abolished by miR-15b-5p mimics. Importantly, the stemness of cancer stem cells was mediated by the Linc00662/miR-15b-5p axis. Conclusion: Patients with HSCC with high Linc00662 showed poor prognosis and high Linc00662 induced stemness of tumor cells by targeting miR-15b-5p. Linc00662 may serve as a novel diagnostic and target marker for head and neck squamous cell carcinoma.
摘要:
背景:长链非编码RNA(lncRNAs)与多种头颈部肿瘤相关,并在癌症中起重要作用。本研究探讨了Linc00662在下咽鳞状细胞癌(HSCC)中的分子机制。方法:采用实时定量逆转录聚合酶链反应(qRT-PCR)检测HSCC组织中基因的表达。使用CCK-8测定法测量肿瘤细胞的活力和增殖。使用流式细胞术和蛋白质印迹法测量HSCC细胞凋亡。使用球体形成测定来检查细胞干细胞性。建立异种移植肿瘤模型以研究Linc00662在体内的作用。结果:Linc00662在HSCC组织中的表达水平明显高于癌旁正常组织。Linc00662的表达与肿瘤分期无显著关系。发现具有高Linc00662表达的患者具有比具有低Linc00662表达的患者更短的总生存期。Linc00662过表达促进细胞活力并抑制细胞凋亡。使用在线数据库和双荧光素酶报告基因,miR-15b-5p被证实为Linc00662的潜在下游海绵。此外,Linc00662在HSCC细胞中与miR-15b-5p呈负相关。miR-15b-5p的耗尽可以在体内和体外逆转Linc00662的功能。此外,Linc00662促进肿瘤生长,它被miR-15b-5p模拟物废除。重要的是,肿瘤干细胞的干性由Linc00662/miR-15b-5p轴介导。结论:高Linc00662的HSCC患者预后差,高Linc00662通过靶向miR-15b-5p诱导肿瘤细胞干性。Linc00662可作为头颈部鳞状细胞癌的新型诊断和靶标志物。