USP14 通过促进 SOX2 去泛素化来促进 OSCC 的癌症干细胞样细胞特性。
USP14 promotes the cancer stem-like cell properties of OSCC via promoting SOX2 deubiquitination.
机构信息
State Key Laboratory of Oral Diseases, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, China.
出版信息
Oral Dis. 2024 Oct;30(7):4255-4265. doi: 10.1111/odi.14896. Epub 2024 Feb 20.
OBJECTIVE
USP14 (Ubiquitin-specific-processing protease 14) is a deubiquitinating enzyme with oncogenic effects in oral squamous cell carcinoma (OSCC). This study aims to identify new substrates of USP14 and elucidate their role in modulating cancer stem-like cells (CSCs) in OSCC.
MATERIALS AND METHODS
Bioinformatics prediction and docking were performed using UbiBrowser 2.0 and HDOCK, respectively. OSCC cell lines and patient-derived cells were used for experimental validation, employing co-immunoprecipitation, cycloheximide chase assays, and tumor sphere formation to evaluate the effects of USP14 on SOX2 stability, ubiquitination, and CSC phenotypes.
RESULTS
USP14 upregulation was associated with worse overall survival and progression-free interval in OSCC. USP14 interacted with SOX2 with its ubiquitin carboxyl-terminal hydrolase domain. USP14 knockdown impaired SOX2 stability by increasing its polyubiquitination. Ectopic overexpression of wild-type USP14, but not the hydrolase-deficient-mutant USP14, enhanced SOX2 stability by reducing polyubiquitination. USP14 knockdown suppressed OSCC cell proliferation, colony formation, and tumor sphere formation in vitro and tumor growth in vivo. However, the reduction of CSC markers following USP14 knockdown was mitigated by overexpressing SOX2. These findings were verified in OSCC patient-derived CSC cells.
CONCLUSION
This study revealed a USP14-SOX2 axis regulating the CSC properties of OSCC.
目的
USP14（泛素特异性加工蛋白酶 14）是一种具有致癌作用的口腔鳞状细胞癌（OSCC）中的去泛素化酶。本研究旨在鉴定 USP14 的新底物，并阐明其在调节 OSCC 中的癌症干细胞样细胞（CSC）中的作用。
材料和方法
使用 UbiBrowser 2.0 和 HDOCK 分别进行生物信息学预测和对接。使用 OSCC 细胞系和患者来源的细胞进行实验验证，采用免疫共沉淀、环己酰亚胺追踪实验和肿瘤球体形成实验来评估 USP14 对 SOX2 稳定性、泛素化和 CSC 表型的影响。
结果
USP14 的上调与 OSCC 患者的总生存期和无进展间隔缩短相关。USP14 与 SOX2 通过其泛素羧基末端水解酶结构域相互作用。USP14 敲低通过增加其多泛素化来破坏 SOX2 的稳定性。野生型 USP14 的过表达而非水解酶缺陷突变体 USP14 的过表达通过减少多泛素化来增强 SOX2 的稳定性。USP14 敲低抑制 OSCC 细胞在体外的增殖、集落形成和肿瘤球体形成以及体内的肿瘤生长。然而，USP14 敲低后 CSC 标志物的减少被过表达 SOX2 减轻。这些发现在 OSCC 患者来源的 CSC 细胞中得到了验证。
结论
本研究揭示了 USP14-SOX2 轴调节 OSCC 的 CSC 特性。
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