Relevance of copper transporter 1 for cisplatin resistance in human ovarian carcinoma cells
Relevance of copper transporter 1 for cisplatin resistance in human ovarian carcinoma cells
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DOI：
10.1016/j.jinorgbio.2012.07.010
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发表时间：
2012-11-01
影响因子：
3.9
通讯作者：
Jaehde, Ulrich
中科院分区：
文献类型：
--
作者：
Kalayda, Ganna V.;Wagner, Christina H.;Jaehde, Ulrich
抗肿瘤药物顺铂的细胞内积累缺陷是在选择顺铂抗性的细胞中常见的特征。铜转运蛋白1（CTR 1）在药物摄取和耐药中起重要作用。在这里，我们描述了一个详细的调查CTR 1参与顺铂的吸收和顺铂耐药的相关性，使用一个良好的特点敏感/顺铂耐药细胞系对：A2780人卵巢癌细胞系及其顺铂耐药变异A2780 cis。与A2780细胞相比，A2780 cis细胞显示顺铂蓄积减少和CTR 1表达降低。与硫酸铜共孵育既不影响顺铂蓄积（通过无名原子吸收光谱法测定），也不影响其细胞毒性（使用MTT测定法测定，MTT=3-（4，5-二甲基噻唑-2-基）-2，5-二苯基-2H-溴化四唑）。在这两种细胞系中，CTR 1定位于细胞核附近，如使用共聚焦荧光显微镜所发现的。蛋白质在核周区域的稳态定位似乎涉及其从细胞表面的持续内吞作用。与铜相反，顺铂暴露对CTR 1的亚细胞定位没有影响。CTR 1和荧光顺铂类似物标记的羧基荧光素-二乙酸酯之间的共定位可以观察到泡状结构时，连续检索的蛋白质从细胞膜被抑制。我们的结果强烈表明，CTR 1介导所研究细胞系中顺铂的吸收。在通过CTR 1转运穿过质膜后，铂药物可能与蛋白质一起被沿着。我们的研究结果表明，减少CTR 1的表达占减少顺铂的积累，并在A2780 cis细胞系顺铂耐药的决定因素之一。(c)2012 Elsevier Inc. All rights reserved.
Defects in intracellular accumulation of the antitumour drug cisplatin are a commonly observed feature in the cells selected for cisplatin resistance. Copper transporter 1 (CTR1) has been suggested to play an important role in drug uptake and resistance. Here, we describe a detailed investigation of the involvement of CTR1 in cisplatin uptake and its relevance for cisplatin resistance using a well characterised sensitive/cisplatin-resistant cell line pair: A2780 human ovarian carcinoma cell line and its cisplatin-resistant variant A2780cis. A2780cis cells showed decreased cisplatin accumulation and lower CTR1 expression compared to A2780 cells. Co-incubation with copper sulphate affected neither cisplatin accumulation (determined by nameless atomic absorption spectrometry) nor its cytotoxicity (determined using an MTT-assay, MTT=3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide). In both cell lines, CTR1 was localised near the nucleus as found using confocal fluorescence microscopy. The steady-state localisation of the protein in perinuclear region appears to involve its continuous endocytosis from cell surface. In contrast to copper, cisplatin exposure had no influence on the sub cellular localisation of CTR1. Co-localisation between CTR1 and a fluorescent cisplatin analogue labelled with carboxyfluorescein-diacetate could be observed in vesicular structures when continuous retrieval of the protein from cell membrane was inhibited. Our results strongly suggest that CTR1 mediates cisplatin uptake in the cell lines studied. Upon its transport across the plasma membrane by CTR1 the platinum drug is likely to be internalised along with the protein. Our findings imply that reduced CTR1 expression accounts for decreased cisplatin accumulation and represents one of the determinants of cisplatin resistance in A2780cis cell line. (c) 2012 Elsevier Inc. All rights reserved.