Mertins Philipp, Yang Feng, Liu Tao, Mani D R, Petyuk Vladislav A, Gillette Michael A, Clauser Karl R, Qiao Jana W, Gritsenko Marina A, Moore Ronald J, Levine Douglas A, Townsend Reid, Erdmann-Gilmore Petra, Snider Jacqueline E, Davies Sherri R, Ruggles Kelly V, Fenyo David, Kitchens R Thomas, Li Shunqiang, Olvera Narciso, Dao Fanny, Rodriguez Henry, Chan Daniel W, Liebler Daniel, White Forest, Rodland Karin D, Mills Gordon B, Smith Richard D, Paulovich Amanda G, Ellis Matthew, Carr Steven A
From the ‡Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142;
‖Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352;
Mol Cell Proteomics. 2014 Jul;13(7):1690-704. doi: 10.1074/mcp.M113.036392. Epub 2014 Apr 9.
Protein abundance and phosphorylation convey important information about pathway activity and molecular pathophysiology in diseases including cancer, providing biological insight, informing drug and diagnostic development, and guiding therapeutic intervention. Analyzed tissues are usually collected without tight regulation or documentation of ischemic time. To evaluate the impact of ischemia, we collected human ovarian tumor and breast cancer xenograft tissue without vascular interruption and performed quantitative proteomics and phosphoproteomics after defined ischemic intervals. Although the global expressed proteome and most of the >25,000 quantified phosphosites were unchanged after 60 min, rapid phosphorylation changes were observed in up to 24% of the phosphoproteome, representing activation of critical cancer pathways related to stress response, transcriptional regulation, and cell death. Both pan-tumor and tissue-specific changes were observed. The demonstrated impact of pre-analytical tissue ischemia on tumor biology mandates caution in interpreting stress-pathway activation in such samples and motivates reexamination of collection protocols for phosphoprotein analysis.
蛋白质丰度和磷酸化传递了有关包括癌症在内的疾病中信号通路活性和分子病理生理学的重要信息，为生物学研究提供见解，为药物和诊断开发提供依据，并指导治疗干预。所分析的组织通常在没有严格控制或记录缺血时间的情况下采集。为了评估缺血的影响，我们在不中断血管的情况下采集了人卵巢肿瘤和乳腺癌异种移植组织，并在设定的缺血时间间隔后进行了定量蛋白质组学和磷酸化蛋白质组学分析。尽管在60分钟后，整体表达的蛋白质组和超过25000个定量磷酸化位点中的大多数没有变化，但在高达24%的磷酸化蛋白质组中观察到了快速的磷酸化变化，这代表了与应激反应、转录调控和细胞死亡相关的关键癌症信号通路的激活。观察到了泛肿瘤和组织特异性的变化。分析前组织缺血对肿瘤生物学的影响表明，在解释此类样本中的应激信号通路激活时需要谨慎，并促使重新审视磷酸化蛋白质分析的采集方案。
