Epidermal growth factor receptor pathway analysis identifies amphiregulin as a key factor for cisplatin resistance of human breast cancer cells
Epidermal growth factor receptor pathway analysis identifies amphiregulin as a key factor for cisplatin resistance of human breast cancer cells
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DOI：
10.1074/jbc.m706287200
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发表时间：
2008-01-11
影响因子：
4.8
通讯作者：
Royer, Hans-Dieter
中科院分区：
文献类型：
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作者：
Eckstein, Niels;Servan, Kati;Royer, Hans-Dieter
使用铂络合物治疗乳腺癌是一种新兴的治疗方式。为了深入了解乳腺癌顺铂耐药的机制，我们使用雌激素受体阳性MCF-7细胞作为模型系统。我们产生了顺铂耐药的MCF-7细胞，并确定了表皮生长因子受体（EGFR），MAPK和AKT信号通路的磷酸化酪氨酸激酶和磷酸化MAPK阵列的功能状态。顺铂耐药MCF-7细胞的特征在于EGFR磷酸化增加、高水平的AKT 1激酶活性和ERK 1磷酸化。与此相反，MAPK信号通路的JNK和p38 MAPK模块无活性。这些条件与p53通路的失活和BCL-2表达增加有关。我们研究了编码ERBB信号级联配体的基因的表达，发现双调蛋白表达的选择性上调发生在顺铂耐药发展的后期。双调蛋白是EGFR（ERBB 1）的特异性配体，并且是上皮细胞的有效促分裂原。暴露于顺铂后，耐药MCF-7细胞分泌的双调蛋白蛋白在较长的时间内，和敲低的双调蛋白表达的特异性短干扰RNA导致几乎完全逆转的耐药表型。为了证明我们的研究结果的普遍性和重要性，我们研究了多种人乳腺癌细胞系中双调蛋白表达和顺铂耐药性，发现了高度显著的相关性。相反，在一组肺癌细胞系中，双调蛋白水平与顺铂耐药性没有显著相关性。因此，我们已经确定了一个新的功能，双调蛋白的顺铂耐药的人乳腺癌细胞。
The use of platinum complexes for the therapy of breast cancer is an emerging new treatment modality. To gain insight into the mechanisms underlying cisplatin resistance in breast cancer, we used estrogen receptor-positive MCF-7 cells as a model system. We generated cisplatin-resistant MCF-7 cells and determined the functional status of epidermal growth factor receptor ( EGFR), MAPK, and AKT signaling pathways by phosphoreceptor tyrosine kinase and phospho-MAPK arrays. The cisplatin-resistant MCF-7 cells are characterized by increased EGFR phosphorylation, high levels of AKT1 kinase activity, and ERK1 phosphorylation. In contrast, the JNK and p38 MAPK modules of the MAPK signaling pathway were inactive. These conditions were associated with inactivation of the p53 pathway and increased BCL-2 expression. We investigated the expression of genes encoding the ligands for the ERBB signaling cascade and found a selective up-regulation of amphiregulin expression, which occurred at later stages of cisplatin resistance development. Amphiregulin is a specific ligand of the EGFR (ERBB1) and a potent mitogen for epithelial cells. After exposure to cisplatin, the resistant MCF-7 cells secreted amphiregulin protein over extended periods of time, and knockdown of amphiregulin expression by specific short interfering RNA resulted in a nearly complete reversion of the resistant phenotype. To demonstrate the generality and importance of our findings, we examined amphiregulin expression and cisplatin resistance in a variety of human breast cancer cell lines and found a highly significant correlation. In contrast, amphiregulin levels did not significantly correlate with cisplatin resistance in a panel of lung cancer cell lines. We have thus identified a novel function of amphiregulin for cisplatin resistance in human breast cancer cells.