Cancer Hospital/Cancer Institute, College of Life Sciences and Institutes of Biomedical Sciences, Fudan University, Shanghai, PR China.
Proteomics. 2010 Apr;10(7):1374-90. doi: 10.1002/pmic.200900380.
The proteins involved in breast cancer initiation and progression are still largely elusive. To gain insights into these processes, we conducted quantitative proteomic analyses with 21T series of breast cell lines, which include a normal, primary tumor and a metastatic tumor that were isolated from a single patient. Stable isotope labeling of amino acid in cell culture followed by LC-MS/MS analysis was performed and deregulated proteins were identified using statistical analysis. Gene ontology analysis revealed that proteins involved in metabolic processes were the most deregulated in both tumorigenesis and metastasis. Interaction network analysis indicated that ERBB2 signaling played a critical role in tumorigenesis. In addition to known markers such as ERBB2 and E-cadherin, novel markers, including BRP44L, MTHFD2 and TIMM17A, were found to be overexpressed in 21T breast cancer cells and verified in additional breast cell lines. mRNA expression analysis as well as immunohistochemistry analysis in breast cancer tissues indicated that expression level of TIMM17A was directly correlated with tumor progression, and survival analysis suggested that TIMM17A was a powerful prognosis factor in breast cancer. More interestingly, overexpression and siRNA knockdown experiments indicated an oncogenic activity of TIMM17A in breast cancer. Our study provides a list of potential novel markers for breast cancer tumorigenesis and metastasis using a unique cell model. Further studies on TIMM17A as well as other markers on the list may reveal mechanisms that result in more effective therapeutics for cancer treatment.
涉及乳腺癌发生和进展的蛋白质仍然很大程度上难以捉摸。为了深入了解这些过程，我们使用 21T 系列乳腺细胞系进行了定量蛋白质组学分析，其中包括来自单个患者的正常、原发性肿瘤和转移性肿瘤。采用稳定同位素标记氨基酸的细胞培养物，然后进行 LC-MS/MS 分析，并使用统计分析鉴定失调蛋白。GO 分析显示，在肿瘤发生和转移过程中，涉及代谢过程的蛋白质失调最为严重。互作网络分析表明 ERBB2 信号在肿瘤发生中起着关键作用。除了 ERBB2 和 E-cadherin 等已知标志物外，还发现 BRP44L、MTHFD2 和 TIMM17A 等新型标志物在 21T 乳腺癌细胞中过度表达，并在其他乳腺细胞系中得到验证。mRNA 表达分析和乳腺癌组织的免疫组织化学分析表明，TIMM17A 的表达水平与肿瘤进展直接相关，生存分析表明 TIMM17A 是乳腺癌的一个强有力的预后因素。更有趣的是，过表达和 siRNA 敲低实验表明 TIMM17A 在乳腺癌中具有致癌活性。我们的研究使用独特的细胞模型为乳腺癌的发生和转移提供了一组潜在的新型标志物。对 TIMM17A 以及列表上的其他标志物的进一步研究可能揭示导致癌症治疗更有效治疗的机制。
