Abstract：
Oral squamous cell carcinoma (OSCC) is a common malignant tumor with high recurrence, metastasis rates, and poor prognosis. Numerous studies discover that circular RNA (circRNA) is closely associated with OSCC progression. Hsa_circ_0020377 has been aberrantly expressed in OSCC, but its role in tumor growth and metastasis remains largely unclear. Hsa_circ_0020377, microRNA-194-5p (miR-194-5p), and Krüppel-like factor 7 (KLF7) contents were determined by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferative, cycle progression migration, and invasion were measured using 5-ethynyl-2\'-deoxyuridine (EdU), Cell Counting Kit-8 (CCK-8), flow cytometry, wound healing, and Transwell assays. The glycolysis level was detected via specific kits. Cyclin D1, E-cadherin, hexokinase 2 (HK2), and KLF7 protein levels were detected via western blot. Using predicting bioinformatics software, the binding between miR-194-5p and hsa_circ_0020377 or KLF7 was verified using a dual-luciferase reporter and RNA Immunoprecipitation (RIP). Beyond that, a xenograft tumor model was used to analyze the role of hsa_circ_0020377 on tumor cell growth in vivo. Increased hsa_circ_0020377 and KLF7 and reduced miR-194-5p were found in OSCC tissues and cell lines. Loss-of-function experiments proved that hsa_circ_0020377 depletion might block OSCC cell proliferation, cycle progression, migration, invasion, and glycolysis in vitro. In xenograft mouse models, hsa_circ_0020377 silencing might suppress tumor growth. In addition, mechanism research suggested that hsa_circ_0020377 could bind with miR-194-5p and enhance its target gene (KLF7), thereby affecting OSCC development. These results broaden our insights regarding the regulation of OSCC progression via circRNA and act as a reference for future clinical studies in OSCC diagnosis and treatment.
摘要：
口腔鳞状细胞癌（OSCC）是一种常见的恶性肿瘤，具有较高的复发率。转移率，预后不良。大量研究发现环状RNA(circularRNA,circRNA)与OSCC进展密切相关。Hsa_circ_0020377在OSCC中异常表达,但其在肿瘤生长和转移中的作用仍不清楚。Hsa_circ_0020377,microRNA-194-5p(miR-194-5p),通过实时定量聚合酶链反应（RT-qPCR）测定Krüppel样因子7（KLF7）的含量。细胞增殖性,循环进程迁移，使用5-乙炔基-2'-脱氧尿苷（EdU）测量侵袭，细胞计数套件-8（CCK-8），流式细胞术,伤口愈合，和Transwell分析。通过特定试剂盒检测糖酵解水平。细胞周期蛋白D1，E-钙粘蛋白，己糖激酶2（HK2），通过蛋白质印迹检测KLF7蛋白水平。使用预测生物信息学软件，miR-194-5p与hsa_circ_0020377或KLF7之间的结合使用双荧光素酶报告基因和RNA免疫沉淀(RIP)进行验证.除此之外，使用异种移植肿瘤模型来分析hsa_circ_0020377对体内肿瘤细胞生长的作用。在OSCC组织和细胞系中发现hsa_circ_0020377和KLF7增加和miR-194-5p减少。功能缺失实验证明，hsa_circ_0020377耗竭可能阻断OSCC细胞增殖，周期进展，迁移,入侵,和体外糖酵解。在异种移植小鼠模型中，hsa_circ_0020377沉默可能抑制肿瘤生长。此外,机制研究表明,hsa_circ_0020377可以与miR-194-5p结合并增强其靶基因(KLF7),从而影响OSCC的发展。这些结果拓宽了我们关于通过circRNA调节OSCC进展的见解，并作为未来OSCC诊断和治疗临床研究的参考。
