International Journal of Oncology, 2016, 48(3).

Quantitative phosphoproteomics reveals genistein as a modulator of cell cycle and DNA damage response pathways in triple-negative breast cancer cells

Yi Fang, Qian Zhang, Xin Wang, et al

Department of Breast Surgical Oncology, Cancer Institute and Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China; et al

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.

《国际肿瘤杂志》2016

定量蛋白质组研究表明在三阴乳腺癌细胞中金雀异黄素是细胞周期和 DNA 损伤反应途径的调节器

方毅，张倩，王新等

中国医学科学院和北京协和医学院肿瘤研究所和医院乳腺外科肿瘤科，北京，中国; et al

大约六分之一的乳腺癌病例被分类为三阴性乳腺癌（TNBC），其在不存在雌激素受体（ER），孕酮受体（PR）和人表皮生长因子受体2（HER2）的表达的情况下命名;然而，TNBC患者的临床结果差和靶向治疗短缺。染料木黄酮，雌激素大豆异黄酮，在TNBC细胞中显示抗癌作用，例如诱导G2/M细胞周期停滞和细胞凋亡。然而，其抗癌作用的基础机制了解很少，其阐释可以帮助TNBC的新型治疗策略的发展。在本研究中，通过结合基于同量异位标签的TMT标记与基于二氧化钛的磷酸肽富集，我们在染料木素处理后定量TNBC细胞系MDA-MB-231中的2,008个磷酸化蛋白上的5,445个磷酸化位点。我们的分析显示在226蛋白质上有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过程，包括DNA复制，凝集素复合物裂解和动粒形成。此外，染料木黄酮还可以激活DNA损伤反应，包括激活ATR和BRCA1复杂。总的来说，我们的研究提出了在磷酸化蛋白水平的证据，染料木黄酮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。