WCU Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology and College of Medicine or College of Pharmacy, Seoul National University, Seoul, South Korea.
Proteomics. 2013 May;13(10-11):1714-25. doi: 10.1002/pmic.201200511. Epub 2013 Apr 23.
Cancer stem cells (CSCs) are a subpopulation of tumor cells that can self-renew, metastasize, and promote cancer recurrence. A comprehensive characterization of the CSC proteome has been hampered due to their scarcity and rapid differentiation. Here, we present a systematic analysis of the cell-surface proteome using a CSC-like cell line derived from MDA-MB453 breast cancer cells, which exhibited a CD44(+) /CD24(-) (where CD is cluster of differentiation) phenotype and chemoresistance. We identified differentially expressed proteins in CSC-like cells, including upregulated plasma membrane proteins such as CD44, CD133, epidermal growth factor receptor (EGFR), CD147, cadherin 1, integrins, and catenin (cadherin-associated protein), beta 1 (CTNNB1), using an in-situ biotinylation approach followed by MS analysis. We examined the role of CD147 in the promotion of CSC growth and survival, and demonstrated that inhibition of CD147 with a monoclonal antibody induced significant inhibition of cell growth. siRNA-mediated silencing of CD147 gene expression restored the sensitivity of CSC-like cells to 5-fluorouracil (5-FU), along with decreasing the expression of thymidylate synthase, p-AKT, and β-catenin, while increasing the expression of p-glycogen synthase kinase (GSK)3β. Increased CD147 expression in the CSC-like cells, as seen by proteomic analysis, and the functional consequences of CD147 overexpression in CSC-like cells suggest that CD147 may be one of the critical cell-surface proteins involved in promoting chemoresistance and survival in CSCs.
癌症干细胞（CSC）是肿瘤细胞中的一个亚群，能够自我更新、转移并促进癌症复发。由于其稀缺性和快速分化，对 CSC 蛋白质组的全面描述一直受到阻碍。在这里，我们使用源自 MDA-MB453 乳腺癌细胞的 CSC 样细胞系对细胞表面蛋白质组进行了系统分析，该细胞系表现出 CD44(+) / CD24(-)（CD 是分化群）表型和化学抗性。我们在 CSC 样细胞中鉴定出差异表达的蛋白质，包括上调的质膜蛋白，如 CD44、CD133、表皮生长因子受体 (EGFR)、CD147、钙粘蛋白 1、整合素和连环蛋白（钙粘蛋白相关蛋白）、β1 (CTNNB1)，使用原位生物素化方法后进行 MS 分析。我们研究了 CD147 在促进 CSC 生长和存活中的作用，并证明用单克隆抗体抑制 CD147 可显著抑制细胞生长。CD147 基因表达的 siRNA 介导沉默恢复了 CSC 样细胞对 5-氟尿嘧啶（5-FU）的敏感性，同时降低了胸苷酸合成酶、p-AKT 和β-连环蛋白的表达，而增加了 p-糖原合酶激酶 (GSK)3β 的表达。CSC 样细胞中 CD147 表达的增加，如蛋白质组分析所示，以及 CD147 过表达在 CSC 样细胞中的功能后果表明，CD147 可能是参与促进 CSCs 化学抗性和存活的关键细胞表面蛋白之一。
