Jain Ankit P, Ghose Vivek, Munshi Srijon, Bhat Firdous A, Dey Gourav, Nanjappa Vishalakshi
Institute of Bioinformatics, International Technology Park, Bangalore, India.
Manipal Academy of Higher Education (MAHE), Manipal, India.
Mol Cell Oncol. 2024 Mar 13;11(1):2328873. doi: 10.1080/23723556.2024.2328873. eCollection 2024.
Cisplatin is the commonly used chemotherapeutic drug in treatment of various cancers. However, development of resistance towards cisplatin results in tumor recurrence. Here, we aim to understand the mechanisms of action of cisplatin and emergence of resistance to cisplatin using mass spectrometry-based proteomic approach. A panel of head and neck squamous cell carcinoma (HNSCC) cell lines were treated with cisplatin at respective IC for 24 h and label-free mass spectrometry analysis was carried out. Proteomic analysis of A253, FaDu, Det562 and CAL27 cell lines upon cisplatin treatment resulted in the identification of 5,060, 4,816, 4,537 and 4,142 proteins, respectively. Bioinformatics analysis of differentially regulated proteins revealed proteins implicated in DNA damage bypass pathway, translation and mRNA splicing to be enriched. Further, proteins associated with cisplatin resistance exhibited alterations following short-term cisplatin exposure. Among these, class III tubullin protein (TUBB3) was found to be upregulated in cisplatin-treated cells compared to untreated cells. Western blot analysis confirmed the elevated expression of TUBB3 in cells treated with cisplatin for 24 h, and also in cisplatin resistant HNSCC cell lines. This study delineates the early signaling events that enable HNSCC cells to counteract the cytotoxic effects of cisplatin and facilitate the development of resistance.
顺铂是治疗各种癌症常用的化疗药物。然而，对顺铂产生耐药性会导致肿瘤复发。在此，我们旨在使用基于质谱的蛋白质组学方法来了解顺铂的作用机制以及对顺铂耐药性的产生。一组头颈部鳞状细胞癌（HNSCC）细胞系分别用各自的半数抑制浓度（IC）的顺铂处理24小时，并进行无标记质谱分析。对顺铂处理后的A253、FaDu、Det562和CAL27细胞系进行蛋白质组学分析，分别鉴定出5060、4816、4537和4142种蛋白质。对差异调节蛋白的生物信息学分析显示，参与DNA损伤旁路途径、翻译和mRNA剪接的蛋白富集。此外，与顺铂耐药相关的蛋白在短期顺铂暴露后发生了改变。其中，与未处理细胞相比，Ⅲ类微管蛋白（TUBB3）在顺铂处理的细胞中被发现上调。蛋白质印迹分析证实，在顺铂处理24小时的细胞以及顺铂耐药的HNSCC细胞系中，TUBB3的表达均升高。本研究描述了使HNSCC细胞能够对抗顺铂的细胞毒性作用并促进耐药性发展的早期信号事件。
