Protein pathway activation mapping reveals molecular networks associated with antiestrogen resistance in breast cancer cell lines
Protein pathway activation mapping reveals molecular networks associated with antiestrogen resistance in breast cancer cell lines
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DOI：
10.1002/ijc.27489
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发表时间：
2012-11-01
影响因子：
6.4
通讯作者：
Dorssers, Lambert C. J.
中科院分区：
文献类型：
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作者：
van Agthoven, Ton;Godinho, Marcia F. E.;Dorssers, Lambert C. J.
以前，我们已经确定了一组乳腺癌抗雌激素抵抗（BCAR）基因。这些基因中的几个具有临床相关性，因为mRNA或蛋白质水平与他莫昔芬抗性或肿瘤侵袭性相关。我们假设BCAR基因诱导的蛋白质信号网络激活状态的变化可能会更好地了解抗雌激素抵抗的机制。关键的信号转导通路的激活或表达的变化进行了分析，使用反相蛋白质微阵列探测与78个已知的作用，在肿瘤发生的信号蛋白的抗体。我们使用了用AKT 1、AKT 2、BCAR 1、BCAR 3、BCAR 4、EGFR、GRB 7、HRAS、HRASv 12或HEF 1转导的ZR-75-1衍生的细胞系和用BCAR 3、BCAR 4或EGFR转导的MCF 7衍生的细胞系。在抗雌激素耐药细胞系中，我们观察到参与细胞增殖和存活的几种途径的磷酸化增加。所有他莫昔芬耐药细胞系均含有高水平的磷酸化AKT及其生化连接底物Forkhead box O 1/3。ERBB 2、ERBB 3和下游调节剂粘着斑激酶和SHC的激活在BCAR 4过表达的细胞中被激活。观察到活化的AMPK α 1、细胞周期蛋白、STAT 5、STAT 6、ERK 1/2和BCL 2的水平存在显著差异。在雌激素依赖和非依赖细胞系的细胞信号网络的比较揭示了生化连接激酶底物标记，包括系统激活的信号通路参与他莫昔芬耐药。我们的研究结果表明，该模型提供了深入了解乳腺癌进展和抗雌激素抵抗的分子和细胞机制。这些知识可能有助于开发新的靶向治疗方法。
Previously, we have identified a panel of breast cancer antiestrogen resistance (BCAR) genes. Several of these genes have clinical relevance because mRNA or protein levels associate with tamoxifen resistance or tumor aggressiveness. We postulated that changes in activation status of protein signaling networks induced by BCAR genes may provide better insight into the mechanisms underlying antiestrogen resistance. Key signal transduction pathways were analyzed for changes in activation or expression using reverse-phase protein microarrays probed with 78 antibodies against signaling proteins with known roles in tumorigenesis. We used ZR-75-1-derived cell lines transduced with AKT1, AKT2, BCAR1, BCAR3, BCAR4, EGFR, GRB7, HRAS, HRASv12 or HEF1 and MCF7-derived cell lines transduced with BCAR3, BCAR4 or EGFR. In the antiestrogen-resistant cell lines, we observed increased phosphorylation of several pathways involved in cell proliferation and survival. All tamoxifen-resistant cell lines contained high levels of phosphorylated AKT and its biochemically linked substrates Forkhead box O1/3. The activation of ERBB2, ERBB3 and the downstream modulators focal adhesion kinase and SHC were activated in cells with overexpression of BCAR4. Remarkable differences were observed for the levels of activated AMPK alpha1, cyclins, STAT5, STAT6, ERK1/2 and BCL2. The comparison of the cell signaling networks in estrogen-dependent and -independent cell lines revealed biochemically linked kinasesubstrate markers that comprised systemically activated signaling pathways involved in tamoxifen resistance. Our results show that this model provides insights into the molecular and cellular mechanisms of breast cancer progression and antiestrogen resistance. This knowledge may help the development of novel targeted treatments.