Identification of 14-3-3σ as a contributor to drug resistance in human breast cancer cells using functional proteomic analysis
Identification of 14-3-3σ as a contributor to drug resistance in human breast cancer cells using functional proteomic analysis
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DOI：
10.1158/0008-5472.can-05-3801
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发表时间：
2006-03-15
期刊：
影响因子：
11.2
通讯作者：
Zhang, JT
中科院分区：
文献类型：
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作者：
Liu, Y;Liu, HL;Zhang, JT
多药耐药（MDR）是肿瘤治疗成功的主要障碍。为了了解MDR的机制，已经建立了许多癌细胞系，并且已经发现了各种耐药机制，例如ATP阻碍盒（ABC）转运蛋白介导的药物外排。以前，从乳腺癌细胞系MCF 7中选择了抗阿霉素的MDR细胞系MCF 7/AdVp 3000，并且认为ABCG 2的过表达在该衍生细胞系中引起MDR。然而，ABCG 2在MCF 7细胞中的异位过表达不能解释所选择的MCF 7/AdVp 3000细胞的极高耐药性水平。我们推测MCF 7/AdVp 3000细胞一定存在阿霉素选择的其他耐药机制。为了验证这一假设，我们比较了MCF 7和MCF 7/AdVp 3000细胞之间的整体蛋白质谱。经过双向凝胶电泳和基质辅助激光解吸/电离飞行时间质谱分析，17个蛋白点与两个细胞系之间的差异水平进行了鉴定。尽管14-3-3 σ、角蛋白18、角蛋白19、ATP合成酶β、蛋白质二硫键异构酶、热休克蛋白27、组织蛋白酶D、磷酸丙糖异构酶、过氧化物氧还蛋白6和电子转移黄素蛋白在MCF 7/AdVp 3000细胞中增加，但nm 23/H1、过氧化物氧还蛋白2、核磷蛋白1/B23和无机焦磷酸酶在MCF 7/AdVp 3000细胞中减少。使用蛋白质印迹法验证了这些蛋白质的差异水平。此外，功能验证显示，升高的14-3-3 σ表达显著有助于在MCF 7/AdVp 3000细胞中观察到的耐药性。因此，我们的结论是，这些蛋白质可能有助于在MCF 7/AdVp 3000细胞中选择的耐药性，并且它们在肿瘤中的表达改变可能导致对化疗的临床耐药性。
Multidrug resistance (MDR) is a major obstacle to successful cancer treatment. To understand the mechanism of MDR, many cancer cell lines have been established, and various mechanisms of resistance, such as ATP-hinding cassette (ABC) transporter-mediated drug efflux, have been discovered. Previously, a MDR cell line MCF7/AdVp3000 was selected from breast cancer cell line MCF7 against Adriamycin, and overexpression of ABCG2 was thought to cause MDR in this derivative cell line. However, ectopic overexpression of ABCG2 in MCF7 cells could not explain the extremely high drug resistance level of the selected MCF7/AdVp3000 cells. We hypothesized that MCF7/AdVp3000 cells must have other resistance mechanisms selected by Adriamycin. To test this hypothesis, we compared the global protein profiles between MCF7 and MCF7/AdVp3000 cells. Following two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis, 17 protein spots with differential levels between the two cell lines were identified. Although 14-3-3 sigma, keratin 18, keratin 19, ATP synthase beta, protein disulfide isomerase, heat shock protein 27, cathepsin D, triose-phosphate isomerase, peroxiredoxin 6, and electron transfer flavoprotein were increased, nm23/H1, peroxiredoxin 2, nucleophosmin 1/B23, and inorganic pyrophosphatase were decreased in MCF7/AdVp3000 cells. The differential levels of these proteins were validated using Western blot. Furthermore, functional validation showed that the elevated 14-3-3 sigma expression contributes considerably to the observed drug resistance in MCF7/AdVp3000 cells. We, thus, conclude that these proteins likely contribute to the resistance selected in the MCF7/AdVp3000 cells, and their altered expression in tumors may cause clinical resistance to chemotherapy.