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环状 hsa_circ_0020377 通过靶向 miR-194-5p 调控 KLF7 促进口腔鳞状细胞癌进展中的肿瘤细胞恶性行为和糖酵解。

Circular hsa_circ_0020377 regulates KLF7 by targeting miR-194-5p to facilitate tumor cell malignant behaviors and glycolysis in oral squamous cell carcinoma progression.

机构信息

Department of Stomatology, The Fourth Hospital of Hebei Medical University, No. 12, JianKang Road, Shijiazhuang, Hebei Province, 050011, People's Republic of China.

Department of Stomatology, Hengshui People's Hospital, Hengshui, Hebei Province, China.

出版信息

Funct Integr Genomics. 2023 Jan 31;23(1):52. doi: 10.1007/s10142-023-00973-w.

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,具有高复发、转移率和预后不良的特点。大量研究发现,环状 RNA(circRNA)与 OSCC 进展密切相关。Hsa_circ_0020377 在 OSCC 中表达异常,但它在肿瘤生长和转移中的作用仍不清楚。通过实时定量聚合酶链反应(RT-qPCR)测定 hsa_circ_0020377、微小 RNA-194-5p(miR-194-5p)和 Kruppel 样因子 7(KLF7)的含量。使用 5-乙炔基-2'-脱氧尿苷(EdU)、细胞计数试剂盒-8(CCK-8)、流式细胞术、划痕愈合和 Transwell 测定法测量细胞增殖、周期进展、迁移和侵袭。通过特定试剂盒检测糖酵解水平。通过 Western blot 检测细胞周期蛋白 D1、E-钙黏蛋白、己糖激酶 2(HK2)和 KLF7 蛋白水平。使用预测生物信息学软件,通过双荧光素酶报告和 RNA 免疫沉淀(RIP)验证 miR-194-5p 与 hsa_circ_0020377 或 KLF7 的结合。此外,还使用异种移植肿瘤模型分析了 hsa_circ_0020377 在体内肿瘤细胞生长中的作用。在 OSCC 组织和细胞系中发现 hsa_circ_0020377 和 KLF7 增加,miR-194-5p 减少。功能丧失实验证明,hsa_circ_0020377 耗竭可能会阻止 OSCC 细胞在体外的增殖、周期进展、迁移、侵袭和糖酵解。在异种移植小鼠模型中,hsa_circ_0020377 沉默可能会抑制肿瘤生长。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对环状 RNA 调节 OSCC 进展的认识,并为 OSCC 诊断和治疗的未来临床研究提供了参考。

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