Department of Internal Medicine, University of Rome Tor Vergata, Rome, Italy.
J Proteome Res. 2011 Feb 4;10(2):416-28. doi: 10.1021/pr100457n. Epub 2010 Dec 29.
Neuroblastoma is one of the most aggressive solid tumors in the childhood. Therapy resistance to anticancer drugs represents the major limitation to the effectiveness of clinical treatment. To better understand the mechanisms underlying cisplatin resistance, we performed a comparative proteomic study of the human neuroblastoma cell line SH-SY5Y and its cisplatin resistant counterpart by both the classical 2-DE electrophoresis coupled to mass spectrometry and the more innovative label-free nLC-MS(E). The differentially expressed proteins were classified by bioinformatic tools according to their biological functions and their involvement in several cellular processes. Moreover, a meta-mining investigation of protein ontologies was also performed on available data from previously published proteomics studies to highlight the modulation of significant cellular pathways, which may regulate the sensitivity of neuroblastoma to cisplatin. In particular, we hypothesized a major role of the transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. Confocal microscopy experiments, enzyme assay, and Western blotting of proteins regulated by Nrf2 provided evidences that this pathway, playing a protective role in normal cells, may represent a potential novel target to control cisplatin resistance in neuroblastoma.
神经母细胞瘤是儿童期最具侵袭性的实体肿瘤之一。对抗癌药物的治疗耐药性是临床治疗效果的主要限制因素。为了更好地理解顺铂耐药的机制，我们通过经典的 2-DE 电泳结合质谱和更创新的无标记 nLC-MS(E)对人神经母细胞瘤细胞系 SH-SY5Y 及其顺铂耐药对应物进行了比较蛋白质组学研究。通过生物信息学工具，根据蛋白质的生物学功能及其在多种细胞过程中的参与情况对差异表达蛋白进行分类。此外，还对来自先前发表的蛋白质组学研究的可用数据进行了蛋白质本体元数据挖掘调查，以突出调节神经母细胞瘤对顺铂敏感性的重要细胞途径的调节。特别是，我们假设转录因子核因子-红细胞 2 相关因子 2 (Nrf2) 途径的主要作用。通过 Nrf2 调节的蛋白质的共聚焦显微镜实验、酶测定和 Western blot 提供了证据，表明该途径在正常细胞中发挥保护作用，可能成为控制神经母细胞瘤顺铂耐药性的潜在新靶点。
