Quantitative phosphoproteomics reveals genistein as a modulator of cell cycle and DNA damage response pathways in triple-negative breast cancer cells
Quantitative phosphoproteomics reveals genistein as a modulator of cell cycle and DNA damage response pathways in triple-negative breast cancer cells
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定量磷酸蛋白质组学揭示金雀花素是三阴性乳腺癌细胞中细胞周期和 DNA 损伤反应途径的调节剂
DOI：
10.3892/ijo.2016.3327
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发表时间：
2016-03-01
影响因子：
5.2
通讯作者：
Wang, Jing
中科院分区：
文献类型：
--
作者：
Fang, Yi;Zhang, Qian;Wang, Jing
大约六分之一的乳腺癌病例被归类为三阴性乳腺癌（TNBC），以雌激素受体（ER）、孕激素受体（PR）和人表皮生长因子受体2（HER 2）的表达缺失而命名;然而，患有TNBC的患者遭受临床结果差和缺乏靶向治疗。染料木黄酮是一种雌激素大豆黄酮，在TNBC细胞中显示出抗癌作用，如诱导G2/M细胞周期阻滞和凋亡。然而，其抗癌作用的潜在机制知之甚少，其阐明可以帮助开发TNBC的新治疗策略。在这项研究中，通过将基于同量异序标签的TMT标记与基于二氧化钛的磷酸肽富集相结合，我们定量了染料木素处理后TNBC细胞系MDA-MB-231中2，008个磷蛋白上的5，445个磷酸化位点。我们的分析揭示了226个蛋白质上的332个染料木黄酮调节的磷酸化位点。我们的数据表明，染料木黄酮可以调节细胞周期中的几个生物学过程，包括DNA复制，cohesin复合物切割和动粒形成。此外，染料木黄酮还可以激活DNA损伤反应，包括ATR和BRCA 1复合物的激活。总的来说，我们的研究在磷酸化蛋白质组学水平上提供了证据，表明染料木素能够通过以更复杂的方式调节细胞周期和DNA损伤反应来抑制TNBC细胞生长。我们的研究结果有助于阐明染料木素在TNBC细胞中发挥其抗癌作用的机制。
Around one sixth of breast cancer cases are classified as triple-negative breast cancer (TNBC), named after the absence of the expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2); however, patients with TNBC suffer from poor clinical outcome and shortage of targeted therapy. Genistein, an estrogenic soy isoflavone, shows anticancer effects in TNBC cells such as inducing G2/M cell cycle arrest and apoptosis. However, the underlying mechanism of its anticancer effects is poorly understood and its elucidation can help the development of novel therapeutic strategies for TNBC. In this study, by combining isobaric tag-based TMT labeling with titanium dioxide-based phosphopeptide enrichment, we quantitated 5,445 phosphorylation sites on 2,008 phosphoproteins in the TNBC cell line MDA-MB-231, upon genistein treatment. Our analysis revealed 332 genistein-regulated phosphorylation sites on 226 proteins. Our data show that genistein can regulate several biological processes during the cell cycle, including DNA replication, cohesin complex cleavage, and kinetochore formation. Furthermore, genistein can also activate DNA damage response, including activation of ATR and BRCA1 complex. Overall, our study presents evidence at a phosphoproteomic level that genistein is able to inhibit TNBC cell growth by regulating the cell cycle and DNA damage response in a more complex manner. Our findings help elucidate the mechanisms through which genistein exerts its anticancer effects in TNBC cells.