Comparative Proteomic Analysis of Paclitaxel Sensitive A2780 Epithelial Ovarian Cancer Cell Line and Its Resistant Counterpart A2780TC1 by 2D-DIGE: The Role of ERp57
Comparative Proteomic Analysis of Paclitaxel Sensitive A2780 Epithelial Ovarian Cancer Cell Line and Its Resistant Counterpart A2780TC1 by 2D-DIGE: The Role of ERp57
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DOI：
10.1021/pr800856b
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发表时间：
2009-04-01
影响因子：
4.4
通讯作者：
Rotilio, Domenico
中科院分区：
文献类型：
--
作者：
Cicchillitti, Lucia;Di Michele, Michela;Rotilio, Domenico
上皮性卵巢癌是妇科癌症死亡的主要原因。尽管对手术和初始化疗反应良好，但化疗耐药的发生是成功治疗的主要障碍。为了更好地了解紫杉醇耐药的生物学机制，我们将基于DIGE结合质谱（MALDI-TOF和LC-MS/MS）的比较蛋白质组学方法应用于人上皮性卵巢癌细胞系A2780及其紫杉醇耐药对应细胞系A2780TC1。两种细胞系之间的差异表达蛋白大部分属于应激反应（29%）、代谢（21%）和细胞周期和凋亡（17%）。我们重点研究了受紫杉醇耐药调节最强烈的蛋白质，特别是二硫异构酶ERp57，它可能代表一种化学耐药生物标志物。ERp57被发现与III类β -微管蛋白（TUBB3）相互作用，参与卵巢癌和其他癌症的紫杉醇耐药。此外，我们证明了该蛋白在细胞骨架中的新定位，并描述了ERp57/TUBB3相互作用也发生在核室中，并与由核蛋白、核磷蛋白、hnRNPK和死亡蛋白形成的多聚体复合物相关。我们的数据表明，ERp57通过与TUBB3的相互作用调节紫杉醇治疗后微管与染色体的附着，在卵巢癌的化疗耐药机制中发挥重要作用。
Epithelial ovarian cancer is the leading cause of gynecological cancer mortality. Despite good response to surgery and initial chemotherapy, chemoresistance occurrence represents a major obstacle to a successful therapy. To better understand biological mechanisms at the basis of paclitaxel resistance, a comparative proteomic approach based on DIGE coupled with mass spectrometry (MALDI-TOF and LC-MS/MS) was applied to the human epithelial ovarian cancer cell lines A2780 and its paclitaxel resistant counterpart A2780TC1. Most of the differentially expressed proteins between the two cell lines belong to the class of stress response (29%), metabolism (21%), and cell cycle and apoptosis (17%). We focused on proteins which were most strongly modulated by paclitaxel resistance and in particular on the disulphide isomerase ERp57, which may represent a chemoresistance biomarker. ERp57 was found to interact with class III beta-tubulin (TUBB3), involved in paclitaxel resistance in ovarian and other cancers. Moreover, we demonstrated a novel localization of this protein in cytoskeleton and described that ERp57/TUBB3 interaction occurs also in the nuclear compartment and in association with a multimeric complex formed by nucleolin, nucleophosmin, hnRNPK, and mortalin. Our data suggest that ERp57 plays an important role in chemoresistance mechanisms in ovarian cancer by modulating the attachment of microtubules to chromosomes following paclitaxel treatment through its interaction with TUBB3.