Ingruber Julia, Dudas Jozsef, Sprung Susanne, Lungu Bianca, Mungenast Felicitas
Department of Otorhinolaryngology and Head and Neck Surgery, Medical University of Innsbruck, Austria and University Hospital of Tyrol, 6020 Innsbruck, Austria.
Inpath GmbH, 6020 Innsbruck, Austria.
Biomedicines. 2022 Oct 5;10(10):2482. doi: 10.3390/biomedicines10102482.
This study aims to investigate the role of partial epithelial to mesenchymal transition (pEMT)-related proteins in modulating Cisplatin resistance in head and neck squamous cell carcinoma (HNSCC). SCC-25 cells were pre-treated with TGF-beta1 followed by transient Krüppel-like Factor 4 (KLF4)-overexpression and Cisplatin treatment. Cell growth, cell morphological changes and cell migration were assessed using Juli BR live cell video-microscopy. In addition, Ki-67 and Slug immunostaining and follow-up image cytometric analysis of primary and recurrent HNSCC tumors were performed to evaluate the proliferation index (PI) and the EMT-like phenotype. We observed that proliferating and Slug-positive tumor cells expand after therapy in HNSCC. Subsequently, protein analysis revealed the stabilization of Slug, upregulation of Vimentin and phospho-p38 (p-p38) in Cisplatin-resistant SCC-25 cells. Moreover, KLF4-overexpression contributed to Cisplatin sensitivity by reduction of Slug at the protein level. This work strongly suggests that an pEMT-like pathway is activated in recurrent and Cisplatin-resistant HNSCC. Finally, stable KLF4-overexpression might sensitize HNSCC tumor cells for Cisplatin treatment.
本研究旨在探讨部分上皮-间质转化（pEMT）相关蛋白在调节头颈部鳞状细胞癌（HNSCC）顺铂耐药中的作用。对SCC-25细胞先用转化生长因子-β1（TGF-β1）预处理，然后进行瞬时Krüppel样因子4（KLF4）过表达及顺铂处理。使用Juli BR活细胞视频显微镜评估细胞生长、细胞形态变化和细胞迁移。此外，对原发性和复发性HNSCC肿瘤进行Ki-67和Slug免疫染色以及后续图像细胞分析，以评估增殖指数（PI）和类似上皮-间质转化（EMT）的表型。我们观察到HNSCC中增殖且Slug阳性的肿瘤细胞在治疗后增多。随后，蛋白质分析显示顺铂耐药的SCC-25细胞中Slug稳定、波形蛋白上调以及磷酸化p38（p-p38）上调。此外，KLF4过表达通过在蛋白质水平降低Slug而提高顺铂敏感性。这项研究强烈表明，在复发性和顺铂耐药的HNSCC中激活了类似pEMT的途径。最后，稳定的KLF4过表达可能使HNSCC肿瘤细胞对顺铂治疗敏感。
