Proteomic profiling of lipid rafts in a human breast cancer model of tumorigenic progression.
Proteomic profiling of lipid rafts in a human breast cancer model of tumorigenic progression.
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DOI：
10.1007/s10585-011-9389-5
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发表时间：
2011-08
影响因子：
4
通讯作者：
Stemmer, Paul M
中科院分区：
文献类型：
--
作者：
Caruso, Joseph A;Stemmer, Paul M
肿瘤生物标志物有助于癌症的早期检测，作为干预的治疗靶点，并作为评估治疗反应的诊断指标。为了鉴定新型人类乳腺癌生物标志物，我们分析了从一系列致瘤潜力不断增加的人类乳腺上皮细胞系中分离的脂筏的蛋白质含量。由于脂筏作为蛋白质相互作用的平台，对几个生物过程至关重要，我们假设，与增殖，侵袭和转移相关的蛋白质的丰度将在高度转化的细胞失调。为此目的，使用了MCF 10A上皮谱系，其包括良性MCF 10A细胞、癌前AT和TG 3B细胞以及恶性CA 1a肿瘤细胞。从每个细胞系中分离耐洗涤剂膜，并使用iTRAQ™试剂和串联质谱法鉴定和相对定量蛋白质。鉴定了57种蛋白质，并且1667种肽鉴定，映射到49种蛋白质，包含足够的信息用于半定量分析。当比较恶性与良性细胞时，我们观察到蛋白质组的一致变化，例如G蛋白含量平均降低5.7倍（n=5），糖基磷脂酰肌醇连接蛋白降低2.7倍（n=7），中间丝增加3.3倍（n=9）。几种鉴定的蛋白质，包括小窝蛋白-1，细丝蛋白A，角蛋白5，6和17，以及波形蛋白，是临床研究中真正的或候选的生物标志物，强调了MCF 10A系列作为模型的有用性，以更好地了解癌症进展的生物学机制。
Tumor biomarkers assist in the early detection of cancer, act as therapeutic targets for intervention, and function as diagnostic indicators for the evaluation of therapeutic responses. To identify novel human breast cancer biomarkers, we have analyzed the protein content of lipid rafts isolated from a series of human mammary epithelial cell lines with increasing tumorigenic potential. Since lipid rafts function as platforms for protein interaction critical to several biological processes, we hypothesized that the abundance of proteins associated with proliferation, invasion and metastasis would be dysregulated in highly transformed cells. For this purpose, the MCF10A epithelial lineage, which include benign MCF10A cells, premalignant AT and TG3B cells, and malignant CA1a tumor cells, was utilized. Detergent-resistant membranes were isolated from each line and proteins were identified and relatively quantitated using iTRAQ™ reagents and tandem mass spectrometry. 57 proteins were identified, and 1667 peptide identifications, mapping to 49 proteins, contained sufficient information for semi-quantitative analysis. When comparing malignant to benign cells, we observed consistent alterations in groups of proteins, such as a 5.7-fold average decrease in G protein content (n=5), 2.7-fold decrease glycosylphosphatidylinositol-linked proteins (n=7) and 3.3-fold increase in intermediate filaments (n=9). Several of the identified proteins, including caveolin-1, filamin A, keratins 5,6 & 17, and vimentin, are bona fide or candidate biomarkers in clinical studies, underscoring the usefulness of the MCF10A series as a model to better understand the biological mechanisms underlying cancer progression.