Differential protein expression profiles in estrogen receptor-positive and -negative breast cancer tissues using label-free quantitative proteomics.
Differential protein expression profiles in estrogen receptor-positive and -negative breast cancer tissues using label-free quantitative proteomics.
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DOI：
10.1177/1947601910365896
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发表时间：
2010-03-01
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影响因子：
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通讯作者：
Han, David K
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作者：
Rezaul, Karim;Thumar, Jay Kumar;Han, David K
确定与雌激素受体(ER)状态相关的蛋白质是更好地理解乳腺癌发生的激素依赖性质的第一步。虽然已经进行了一些基因表达分析，但到目前为止还没有系统地研究蛋白质补体。因为蛋白质是治疗药物的主要靶点，在这项研究中，我们试图识别区分ER阳性和阴性乳腺癌的蛋白质组特征。利用高度浓缩的乳腺肿瘤细胞，对3个ERα+和3个ERα-人乳腺肿瘤进行重复分析，鉴定出2,995个具有≥2肽的独特蛋白质。其中，在ERα+和ERα-乳腺癌组织中大量表达的受体酪氨酸激酶和细胞内酪氨酸激酶被鉴定出来。此外，无标记定量蛋白质组分析显示，236个蛋白质在ERpha+和ERpha-乳腺肿瘤中差异表达。其中，141个蛋白质在ERα+中选择性上调，95个蛋白质在ERα-乳腺肿瘤中选择性上调。将差异表达的蛋白质与乳腺癌数据库进行比较，发现其中98个蛋白质之前已被报道与乳腺癌有关。基因本体论显示，脱氢酶、还原酶、细胞骨架蛋白、细胞外基质、水解酶和裂解酶类在ERα+中显著丰富，而选择性钙结合蛋白、膜运输蛋白和细胞骨架蛋白在ERα乳腺肿瘤中显著丰富。生物学过程和途径分析表明，参与氨基酸代谢、蛋白酶体和脂肪酸代谢的蛋白质过度表达ERpha+上调的蛋白质，而参与糖酵解途径的蛋白质过度表达ERpha-上调的蛋白质。用免疫组织化学方法对4种差异丰富的蛋白质(liprin-alpha1、Fascin、DAP5和β-arrestin-1)的存在和相对丰度进行了定量和验证。综上所述，与体外细胞培养模型不同，我们直接从人乳腺癌组织中鉴定的体内信号蛋白和信号通路可能成为乳腺癌药物干预的相关治疗靶点。
Identification of the proteins that are associated with estrogen receptor (ER) status is a first step towards better understanding of the hormone-dependent nature of breast carcinogenesis. Although a number of gene expression analyses have been conducted, protein complement has not been systematically investigated to date. Because proteins are primary targets of therapeutic drugs, in this study, we have attempted to identify proteomic signatures that demarcate ER-positive and -negative breast cancers. Using highly enriched breast tumor cells, replicate analyses from 3 ERalpha+ and 3 ERalpha- human breast tumors resulted in the identification of 2,995 unique proteins with ≥2 peptides. Among these, a number of receptor tyrosine kinases and intracellular kinases that are abundantly expressed in ERalpha+ and ERalpha- breast cancer tissues were identified. Further, label-free quantitative proteome analysis revealed that 236 proteins were differentially expressed in ERalpha+ and ERalpha- breast tumors. Among these, 141 proteins were selectively up-regulated in ERalpha+, and 95 proteins were selectively up-regulated in ERalpha- breast tumors. Comparison of differentially expressed proteins with a breast cancer database revealed 98 among these have been previously reported to be involved in breast cancer. By Gene Ontology molecular function, dehydrogenase, reductase, cytoskeletal proteins, extracellular matrix, hydrolase, and lyase categories were significantly enriched in ERalpha+, whereas selected calcium-binding protein, membrane traffic protein, and cytoskeletal protein were enriched in ERalpha- breast tumors. Biological process and pathway analysis revealed that up-regulated proteins of ERalpha+ were overrepresented by proteins involved in amino acid metabolism, proteasome, and fatty acid metabolism, while up-regulated proteins of ERalpha- were overrepresented by proteins involved in glycolysis pathway. The presence and relative abundance of 4 selected differentially abundant proteins (liprin-alpha1, fascin, DAP5, and beta-arrestin-1) were quantified and validated by immunohistochemistry. In conclusion, unlike in vitro cell culture models, the in vivo signaling proteins and pathways that we have identified directly from human breast cancer tissues may serve as relevant therapeutic targets for the pharmacological intervention of breast cancer.