TC_global proteomic

<p>The aim of this study was to characterize proteomic and phosphoproteomic alterations in testicular cancer, focusing on two major histopathological subtypes: seminoma and non-seminoma. The analysis also aimed to assess the impact of protein phosphorylation changes between tumor and matched control tissue, as well as between tumor subtypes, on cellular homeostasis and dysregulated biochemical processes potentially associated with early metastatic behavior.</p><p>Tumor and matched control tissues were collected from 46 patients undergoing orchiectomy at the Department of Urology, Medical University of Białystok. Tumor samples were obtained directly from the tumor mass, while control tissue was collected from the contralateral pole of the same testis. Proteomic and phosphoproteomic analyses were performed using an Orbitrap Astral mass spectrometer coupled with nano-flow liquid chromatography. Phosphopeptide enrichment was conducted using Ti IMAC HP and Zn IMAC HP magnetic beads. Data were acquired in Data Independent Acquisition (DIA) mode.</p><p>Global proteomic profiling identified approximately 7,500 proteins, including around 2,500 proteins significantly differentiating tumor tissue from control tissue and approximately 1,800 proteins distinguishing seminoma from non-seminoma. Phosphoproteomic analysis enabled the identification of approximately 3,500 phosphorylated proteins.</p><p>Seminomas were characterized by increased expression of proteins involved in antigen processing and presentation, DNA replication and repair, ribosome biogenesis, cell cycle progression, protein synthesis and folding, and mitochondrial translation. In contrast, seminomas exhibited reduced activity of glutathione metabolism, fatty acid oxidation, and cholesterol biosynthesis. Non-seminomatous tumors displayed enhanced activation of extracellular matrix organization, collagen synthesis, cytoskeletal remodeling, cell adhesion, migration, complement system activation, and endoplasmic reticulum stress response. Although both tumor subtypes showed decreased glutathione metabolism relative to control tissue, non-seminomas demonstrated higher glutathione-related activity compared to seminomas.</p><p>These findings provide comprehensive insights into subtype-specific molecular mechanisms of testicular cancer and identify biological processes potentially contributing to tumor progression and early metastasis.</p>

本研究旨在表征睾丸癌中的蛋白质组学与磷酸化蛋白质组学改变，重点聚焦两大主要组织病理学亚型：精原细胞瘤（seminoma）与非精原细胞瘤（non-seminoma）。本分析还旨在评估肿瘤与配对对照组织之间、以及不同肿瘤亚型之间的蛋白质磷酸化变化，对细胞稳态及潜在与早期转移行为相关的失调生化过程的影响。
研究样本取自比亚韦斯托克医科大学泌尿外科46例行睾丸切除术的患者的肿瘤与配对对照组织。肿瘤样本直接取自肿瘤病灶，对照组织则取自同侧睾丸的对侧极。采用轨道阱Astral（Orbitrap Astral）质谱仪联用纳流液相色谱法开展蛋白质组学与磷酸化蛋白质组学分析。使用钛离子固定金属亲和色谱磁珠（Ti IMAC HP）与锌离子固定金属亲和色谱磁珠（Zn IMAC HP）进行磷酸肽富集。实验数据采用数据非依赖采集（DIA）模式获取。
全局蛋白质组学分析共鉴定得到约7500种蛋白质，其中约2500种可显著区分肿瘤组织与对照组织，约1800种可区分精原细胞瘤与非精原细胞瘤。磷酸化蛋白质组学分析则鉴定得到约3500种磷酸化蛋白质。
精原细胞瘤的特征为参与抗原加工与呈递、DNA复制与修复、核糖体生物发生、细胞周期进程、蛋白质合成与折叠以及线粒体翻译的蛋白质表达上调。与之相反，精原细胞瘤的谷胱甘肽代谢、脂肪酸氧化及胆固醇生物合成通路活性下调。非精原细胞瘤则表现出细胞外基质组织、胶原蛋白合成、细胞骨架重塑、细胞黏附与迁移、补体系统激活以及内质网应激反应通路的活化增强。尽管两种肿瘤亚型相较于对照组织均呈现谷胱甘肽代谢下调，但非精原细胞瘤的谷胱甘肽相关活性高于精原细胞瘤。
本研究结果为睾丸癌的亚型特异性分子机制提供了全面的见解，并鉴定出潜在参与肿瘤进展与早期转移的生物学过程。