Giannopoulou Aikaterini F, Velentzas Athanassios D, Anagnostopoulos Athanasios K, Agalou Adamantia, Papandreou Nikos C, Katarachia Stamatia A, Koumoundourou Dimitra G, Konstantakou Eumorphia G, Pantazopoulou Vasiliki I, Delis Anastasios, Michailidi Maria T, Valakos Dimitrios, Chatzopoulos Dimitris, Syntichaki Popi, Iconomidou Vassiliki A, Tsitsilonis Ourania E, Papassideri Issidora S, Voutsinas Gerassimos E, Hatzopoulos Polydefkis, Thanos Dimitris, Beis Dimitris, Anastasiadou Ema, Tsangaris George Th, Stravopodis Dimitrios J
Section of Cell Biology and Biophysics, Department of Biology, School of Science, National and Kapodistrian University of Athens (NKUA), 15701 Athens, Greece.
Systems Biology Center, Biomedical Research Foundation of the Academy of Athens (BRFAA), 11527 Athens, Greece.
Cancers (Basel). 2021 Apr 22;13(9):2024. doi: 10.3390/cancers13092024.
Melanoma is classified among the most notoriously aggressive human cancers. Despite the recent progress, due to its propensity for metastasis and resistance to therapy, novel biomarkers and oncogenic molecular drivers need to be promptly identified for metastatic melanoma. Hence, by employing nano liquid chromatography-tandem mass spectrometry deep proteomics technology, advanced bioinformatics algorithms, immunofluorescence, western blotting, wound healing protocols, molecular modeling programs, and MTT assays, we comparatively examined the respective proteomic contents of WM115 primary ( = 3955 proteins) and WM266-4 metastatic ( = 6681 proteins) melanoma cells. It proved that WM115 and WM266-4 cells have engaged hybrid epithelial-to-mesenchymal transition/mesenchymal-to-epithelial transition states, with TGF-β controlling their motility in vitro. They are characterized by different signatures of SOX-dependent neural crest-like stemness and distinct architectures of the cytoskeleton network. Multiple signaling pathways have already been activated from the primary melanoma stage, whereas HIF1α, the major hypoxia-inducible factor, can be exclusively observed in metastatic melanoma cells. Invasion-metastasis cascade-specific sub-routines of activated Caspase-3-triggered apoptosis and LC3B-II-dependent constitutive autophagy were also unveiled. Importantly, WM115 and WM266-4 cells exhibited diverse drug response profiles, with epirubicin holding considerable promise as a beneficial drug for metastatic melanoma clinical management. It is the proteome navigation that enables systemic biomarkering and targeted drugging to open new therapeutic windows for advanced disease.
黑色素瘤是最具侵袭性的人类癌症之一。尽管最近取得了进展，但由于其易于转移和对治疗产生抗性，仍需要迅速识别转移性黑色素瘤的新型生物标志物和致癌分子驱动因素。因此，我们采用纳米液相色谱-串联质谱深度蛋白质组学技术、先进的生物信息学算法、免疫荧光、蛋白质印迹、伤口愈合实验方案、分子建模程序和MTT分析，比较研究了WM115原发性（=3955种蛋白质）和WM266-4转移性（=6681种蛋白质）黑色素瘤细胞各自的蛋白质组内容。结果证明，WM115和WM266-4细胞处于混合的上皮-间质转化/间质-上皮转化状态，转化生长因子-β（TGF-β）在体外控制它们的迁移。它们具有不同的SOX依赖性神经嵴样干性特征和独特的细胞骨架网络结构。从原发性黑色素瘤阶段起，多种信号通路就已被激活，而主要的缺氧诱导因子HIF1α仅在转移性黑色素瘤细胞中被观察到。还揭示了活化的半胱天冬酶-3触发的凋亡和LC3B-II依赖性组成性自噬的侵袭-转移级联特异性子程序。重要的是，WM115和WM266-4细胞表现出不同的药物反应谱，表柔比星作为转移性黑色素瘤临床治疗的有益药物具有很大潜力。正是蛋白质组导航使得系统性生物标志物鉴定和靶向给药能够为晚期疾病打开新的治疗窗口。
