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通过对人卵巢癌细胞及其铂耐药亚系进行蛋白质组学分析来鉴定铂耐药相关蛋白。

Identification of platinum-resistance associated proteins through proteomic analysis of human ovarian cancer cells and their platinum-resistant sublines.

作者信息

Yan Xue-dong, Pan Ling-ya, Yuan Ye, Lang Jing-he, Mao Ning

机构信息

Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Shuai Fu Yuan, Beijing 100730, P. R. China.

出版信息

J Proteome Res. 2007 Feb;6(2):772-80. doi: 10.1021/pr060402r.

Abstract

Chemoresistance is a major therapeutic obstacle in cancer patients, and the mechanisms of drug resistance are not fully understood. In the present study, we established platinum-resistant human ovarian cancer cell lines and identified differentially expressed proteins related to platinum resistance. The total proteins of two sensitive (SKOV3 and A2780) and four resistant (SKOV3/CDDP, SKOV3/CBP, A2780/CDDP, and A2780/CBP) human ovarian cancer cell lines were isolated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were identified using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). In total, 57 differential protein spots were identified, and five proteins, including annexin A3, destrin, cofilin 1, Glutathione-S-transferase omega 1 (GSTO1-1), and cytosolic NADP+-dependent isocitrate dehydrogenase (IDHc), were found to be co-instantaneous significance compared with their parental cells. The expression of the five proteins was validated by quantitative PCR and western blot, and the western blot results showed complete consistency with proteomic techniques. The five proteins are hopeful to become candidates for platinum resistance. These may be useful for further study of resistance mechanisms and screening of resistant biomarkers.

摘要

化疗耐药是癌症患者面临的主要治疗障碍,而耐药机制尚未完全明确。在本研究中,我们建立了铂耐药的人卵巢癌细胞系,并鉴定了与铂耐药相关的差异表达蛋白。通过二维凝胶电泳(2-DE)分离了两种敏感的(SKOV3和A2780)和四种耐药的(SKOV3/CDDP、SKOV3/CBP、A2780/CDDP和A2780/CBP)人卵巢癌细胞系的总蛋白。使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定差异表达蛋白。总共鉴定出57个差异蛋白点,发现包括膜联蛋白A3、肌动蛋白解聚因子、丝切蛋白1、谷胱甘肽-S-转移酶ω1(GSTO1-1)和胞质NADP+依赖性异柠檬酸脱氢酶(IDHc)在内的五种蛋白与其亲本细胞相比具有共同的即时显著性。通过定量PCR和蛋白质印迹法验证了这五种蛋白的表达,蛋白质印迹结果与蛋白质组学技术完全一致。这五种蛋白有望成为铂耐药的候选标志物。这些可能有助于进一步研究耐药机制和筛选耐药生物标志物。

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