Proteomic Profiling of Endothelial Cells Exposed to Mitomycin C

Genotoxic stress in mammalian cells defined as a situation that initiates DNA damage compromising the cell’s genomic integrity leading to replication and transcription arrest underlies many pathological conditions including cellular senescence, cancer and cardiovascular diseases. Recent experimental data suggest that genotoxic stress in vitro induced by alkylating mutagen mitomycin C (MMC) is associated with proinflammatory activation of primary human endothelial cells and endothelial-to-mesenchymal transition, the key pathways underlying endothelial disfunction – an initial stage of atherosclerosis, a leading cause of cardiovascular morbidity and mortality worldwide. Given the increasing genotoxic load on the human organism from various environmental (ionizing and UV radiation) and anthropogenic (tobacco smoke, exhaust gases, industrial waste) sources, the decryption of molecular pathways underlying genotoxic stress induced endothelial dysfunction could improve our understanding of atherogenesis and help to justification of genotoxic stress as a novel risk factor for atherosclerosis. Therefore, we performed label-free proteomic profiling of Commercially available primary human coronary artery endothelial cells (HCAEC) and ) and internal thoracic artery endothelial cells (HITAEC) in vitro exposed to MMC followed by bioinformatic analysis to identify biochemical pathways and functional proteins underlying genotoxic stress induced endothelial dysfunction.

哺乳动物细胞中的遗传毒性应激（genotoxic stress），指可引发DNA损伤、破坏细胞基因组完整性，进而导致复制与转录停滞的状态，是细胞衰老、癌症及心血管疾病等诸多病理状态的核心诱因。近期实验数据表明，由烷化剂诱变剂丝裂霉素C（mitomycin C, MMC）在体外诱导的遗传毒性应激，与原代人内皮细胞的促炎活化及内皮细胞向间充质转化（endothelial-to-mesenchymal transition）密切相关；而这两条通路正是内皮功能障碍的关键分子基础——内皮功能障碍是动脉粥样硬化（atherosclerosis）的初始阶段，而动脉粥样硬化是全球范围内引发心血管疾病发病率与死亡率升高的首要病因。鉴于人类机体所承受的遗传毒性负荷日益增加，其来源涵盖各类环境因素（电离辐射与紫外线辐射）及人为活动源（烟草烟雾、汽车尾气、工业废弃物），解析遗传毒性应激诱导内皮功能障碍的分子通路，可加深我们对动脉粥样硬化发生发展的认知，并有助于确立遗传毒性应激作为动脉粥样硬化新型风险因子的地位。因此，本研究对市售原代人冠状动脉内皮细胞（HCAEC）与原代人胸廓内动脉内皮细胞（HITAEC）开展了无标记蛋白质组学分析（label-free proteomic profiling），上述细胞均在体外接受MMC处理，随后通过生物信息学分析，鉴定出与遗传毒性应激诱导内皮功能障碍相关的生化通路与功能蛋白。