Liu Shao-Cheng, Wu Yang-Che, Huang Chih-Ming, Hsieh Ming-Shou, Huang Ting-Yi, Huang Chin-Sheng, Hsu Tung-Nien, Huang Mao-Suan, Lee Wei-Hwa, Yeh Chi-Tai, Lin Chun-Shu
Department of Otolaryngology-Head and Neck Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei City, 114, Taiwan.
School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei City, 110, Taiwan.
Oncogenesis. 2021 Feb 27;10(2):20. doi: 10.1038/s41389-021-00308-z.
Locally advanced oral squamous cell carcinoma (OSCC) requires multimodal therapy, including surgery and concurrent chemoradiotherapy (CCRT). CCRT-resistant and recurrent cancer has a poor prognosis. We investigated the effects of Bruton's tyrosine kinase (BTK) on CCRT-resistant OSCC tissues. The effect of ibrutinib, a first-in-class BTK inhibitor, was tested on stem cell-like OSCC tumorspheres. A tissue array was constructed using tissue samples from 70 patients with OSCC. Human OSCC cell lines, SAS, TW2.6 and HSC-3, were examined. Wound healing, Matrigel invasion, and tumorsphere formation assays, as well as immunofluorescence analysis and flow cytometry, were used to investigate the effects of BTK knockdown (shBTK), ibrutinib, cisplatin, and ibrutinib/cisplatin combination on OSCC cells. We demonstrated that BTK was aberrantly highly expressed in the clinical CCRT-resistant OSCC tissue array, which resulted in poor overall survival in our local Tri-Service General Hospital and freely accessible TCGA OSCC cohorts. shBTK significantly downregulated the stemness markers Nanog, CD133, T cell immunoglobulin-3 (TIM-3), and Krüppel-like factor 4 (KLF4) in SAS tumorspheres and attenuated OSCC cell migration and colony formation. Ibrutinib reduced the number of aldehyde dehydrogenase (ALDH)-rich OSCC cells and reduced tumorsphere formation, migration, and invasion in a dose-dependent manner. Compared with ibrutinib or cisplatin monotherapy, the ibrutinib/cisplatin combination significantly reduced the formation of ALDH + OSCC tumorspheres and enhanced apoptosis. These results demonstrate that ibrutinib effectively inhibits the CSCs-like phenotype of OSCC cells through dysregulation of BTK/CD133 signaling. The ibrutinib/cisplatin combination may be considered for future clinical use.
局部晚期口腔鳞状细胞癌（OSCC）需要多模式治疗，包括手术和同步放化疗（CCRT）。对CCRT耐药和复发的癌症预后较差。我们研究了布鲁顿酪氨酸激酶（BTK）对CCRT耐药OSCC组织的影响。对一流的BTK抑制剂依鲁替尼对干细胞样OSCC肿瘤球的作用进行了测试。使用70例OSCC患者的组织样本构建了组织芯片。检测了人OSCC细胞系SAS、TW2.6和HSC-3。采用伤口愈合、基质胶侵袭和肿瘤球形成试验，以及免疫荧光分析和流式细胞术，研究BTK敲低（shBTK）、依鲁替尼、顺铂和依鲁替尼/顺铂联合用药对OSCC细胞的影响。我们证明，BTK在临床CCRT耐药OSCC组织芯片中异常高表达，这导致我们当地三军总医院和可免费获取的TCGA OSCC队列中的总生存期较差。shBTK显著下调SAS肿瘤球中干性标志物Nanog、CD133、T细胞免疫球蛋白3（TIM-3）和Krüppel样因子4（KLF4），并减弱OSCC细胞迁移和集落形成。依鲁替尼以剂量依赖的方式减少富含醛脱氢酶（ALDH）的OSCC细胞数量，并减少肿瘤球形成、迁移和侵袭。与依鲁替尼或顺铂单药治疗相比，依鲁替尼/顺铂联合用药显著减少ALDH+OSCC肿瘤球形成并增强细胞凋亡。这些结果表明，依鲁替尼通过失调BTK/CD133信号通路有效抑制OSCC细胞的CSCs样表型。依鲁替尼/顺铂联合用药可能值得未来临床应用。
