尽管对于BRAFV600E突变转移性结直肠癌（mCRC）的靶向治疗取得了进展，但BRAFV600E抑制的耐药性发展限制了治疗的响应率和持久性。进一步了解BRAF抑制剂的耐药机制将有助于设计针对BRAF突变的mCRC的新型药理学策略。本研究旨在利用综合蛋白质组学方法，确定与BRAFV600E抑制剂维珀替尼（vemurafenib）耐药相关的新的蛋白质候选物。对所得蛋白质组学数据进行生物信息学分析表明，肌动蛋白细胞骨架连接蛋白（ezrin）作为与维珀替尼耐药显著相关的高排名蛋白质，其在耐药细胞中的过表达在基因和蛋白质水平上得到了进一步证实。NSC305787抑制ezrin通过降低CD44的表达和抑制AKT/c-Myc的活性，增加了耐药细胞对维珀替尼的抗增殖和促凋亡效应，表现为叠加作用效应。我们还发现，在携带BRAFV600E突变的维珀替尼耐药黑色素瘤细胞中，ezrin的表达增加。重要的是，ezrin抑制剂在耐药性黑色素瘤细胞中协同增强了维珀替尼的抗增殖和促凋亡效应。总的来说，我们的研究表明ezrin在携带BRAFV600E突变的结直肠癌和黑色素瘤细胞中对维珀替尼的获得性耐药发挥了作用，并支持进一步的临床前和临床研究，探索联合BRAF抑制剂和肌动蛋白靶向药物作为潜在治疗BRAFV600E突变癌症的方法的益处。

Despite the advancements in targeted therapy for BRAFV600E-mutated metastatic colorectal cancer (mCRC), the development of resistance to BRAFV600E inhibition limits the response rate and durability of the treatment. Better understanding of the resistance mechanisms to BRAF inhibitors will facilitate the design of novel pharmacological strategies for BRAF-mutated mCRC. The aim of this study was to identify novel protein candidates involved in acquired resistance to BRAFV600E inhibitor vemurafenib in BRAFV600E-mutated colon cancer cells using an integrated proteomics approach. Bioinformatic analysis of obtained proteomics data indicated actin-cytoskeleton linker protein ezrin as a highly ranked protein significantly associated with vemurafenib resistance whose overexpression in the resistant cells was additionally confirmed at the gene and protein level. Ezrin inhibition by NSC305787 increased anti-proliferative and pro-apoptotic effects of vemurafenib in the resistant cells in an additive manner, which was accompanied by downregulation of CD44 expression and inhibition of AKT/c-Myc activities. We also detected an increased ezrin expression in vemurafenib-resistant melanoma cells harbouring the BRAFV600E mutation. Importantly, ezrin inhibition potentiated anti-proliferative and pro-apoptotic effects of vemurafenib in the resistant melanoma cells in a synergistic manner. Altogether, our study suggests a role of ezrin in acquired resistance to vemurafenib in colon cancer and melanoma cells carrying the BRAFV600E mutation and supports further pre-clinical and clinical studies to explore the benefits of combined BRAF inhibitors and actin-targeting drugs as a potential therapeutic approach for BRAFV600E-mutated cancers.