Integrated Quantitative Analysis of the Phosphoproteome and Transcriptome in Tamoxifen-resistant Breast Cancer
Integrated Quantitative Analysis of the Phosphoproteome and Transcriptome in Tamoxifen-resistant Breast Cancer
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DOI：
10.1074/jbc.m110.156877
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发表时间：
2011-01-07
影响因子：
4.8
通讯作者：
Okada-Hatakeyama, Mariko
中科院分区：
文献类型：
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作者：
Oyama, Masaaki;Nagashima, Takeshi;Okada-Hatakeyama, Mariko
为了从系统水平上了解三苯氧胺耐药的机制，对配体刺激的人乳腺癌细胞MCF-7进行了定量的磷酸蛋白质组和转录组分析。磷酸化蛋白质组数据显示，在配体刺激后，WT细胞比他莫昔芬耐药细胞更富含磷酸化蛋白。令人惊讶的是，配体扰动后磷酸化降低比磷酸化增加更常见。特别是，17β-雌二醇以非常高的速度诱导WT细胞表达下调。17β-雌二醇和ErbB配体Hereglin在WT细胞中诱导几乎相同数量的上调的磷酸化蛋白。利用转录组数据进行的途径和基序活性分析还表明，GSK3β(糖原合成酶3β)和MAPK1/3信号的非调控激活可能与他莫昔芬耐药细胞中cAMP反应元件结合蛋白和AP-1转录因子的激活改变有关，这一假设得到了报告分析的验证。对临床样本的检查发现，在接受他莫昔芬治疗并最终复发的乳腺癌患者中，丝氨酸9处GSK3β的抑制磷酸化显著降低，这意味着GSK3β的激活可能与他莫昔芬耐药表型有关。因此，结合磷蛋白质组和转录组数据集分析揭示了肿瘤细胞中不同的信号转录程序，并为了解他莫昔芬耐药性提供了一个新的分子靶点。
Quantitative phosphoproteome and transcriptome analysis of ligand-stimulated MCF-7 human breast cancer cells was performed to understand the mechanisms of tamoxifen resistance at a system level. Phosphoproteome data revealed that WT cells were more enriched with phospho-proteins than tamoxifen-resistant cells after stimulation with ligands. Surprisingly, decreased phosphorylation after ligand perturbation was more common than increased phosphorylation. In particular, 17 beta-estradiol induced down-regulation in WT cells at a very high rate. 17 beta-Estradiol and the ErbB ligand heregulin induced almost equal numbers of up-regulated phospho-proteins in WT cells. Pathway and motif activity analyses using transcriptome data additionally suggested that deregulated activation of GSK3 beta (glycogen-synthase kinase 3 beta) and MAPK1/3 signaling might be associated with altered activation of cAMP-responsive element-binding protein and AP-1 transcription factors in tamoxifen-resistant cells, and this hypothesis was validated by reporter assays. An examination of clinical samples revealed that inhibitory phosphorylation of GSK3 beta at serine 9 was significantly lower in tamoxifen-treated breast cancer patients that eventually had relapses, implying that activation of GSK3 beta may be associated with the tamoxifen-resistant phenotype. Thus, the combined phosphoproteome and transcriptome data set analyses revealed distinct signal transcription programs in tumor cells and provided a novel molecular target to understand tamoxifen resistance.