Altered localisation of the copper efflux transporters ATP7A and ATP7B associated with cisplatin resistance in human ovarian carcinoma cells.
Altered localisation of the copper efflux transporters ATP7A and ATP7B associated with cisplatin resistance in human ovarian carcinoma cells.
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DOI：
10.1186/1471-2407-8-175
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发表时间：
2008-06-19
期刊：
影响因子：
3.8
通讯作者：
Jaehde U
中科院分区：
文献类型：
--
作者：
Kalayda GV;Wagner CH;Buss I;Reedijk J;Jaehde U
铜稳态蛋白ATP 7A和ATP 7 B被认为参与顺铂的细胞内转运。本研究的目的是评估这些转运蛋白的亚细胞定位与体外获得性顺铂耐药的相关性。为此，研究了A2780人卵巢癌细胞及其顺铂耐药变异体A2780 cis中ATP 7A和ATP 7 B的定位。在免疫组化染色后，使用共聚焦荧光显微镜研究敏感和耐药细胞中ATP 7A和ATP 7 B的亚细胞定位。进行了与用羧基荧光素-二乙酸酯残基修饰的顺铂类似物的共定位实验。使用基于MTT的测定法测定荧光顺铂类似物在A2780和A2780 cis细胞中的细胞毒性。适当时使用Student t检验或Mann-Whitney检验分析差异的显著性，认为p值< 0.05是显著的。在敏感细胞中，这两种转运蛋白主要定位于高尔基体网络中，而在抗性细胞中，它们被隔离在更外围的囊泡中。A2780 cis细胞中ATP 7A和ATP 7 B的改变定位可能是该细胞系中与溶酶体区室减少相关的细胞内蛋白质运输重大异常的结果。ATP 7A和ATP 7 B亚细胞定位的变化可能有助于顺铂在A2780 cis细胞的囊泡结构中的隔离，这可能会阻止药物与基因组DNA结合，从而导致顺铂耐药性。我们的研究结果表明，转运蛋白的亚细胞定位的改变可能有助于体外顺铂耐药。研究原发性肿瘤细胞培养物和肿瘤组织中的细胞内蛋白定位可能有助于开发临床相关顺铂耐药的标志物。在患者中检测耐药肿瘤反过来可以在治疗的早期阶段实现化疗的个体化。
Copper homeostasis proteins ATP7A and ATP7B are assumed to be involved in the intracellular transport of cisplatin. The aim of the present study was to assess the relevance of sub cellular localisation of these transporters for acquired cisplatin resistance in vitro. For this purpose, localisation of ATP7A and ATP7B in A2780 human ovarian carcinoma cells and their cisplatin-resistant variant, A2780cis, was investigated. Sub cellular localisation of ATP7A and ATP7B in sensitive and resistant cells was investigated using confocal fluorescence microscopy after immunohistochemical staining. Co-localisation experiments with a cisplatin analogue modified with a carboxyfluorescein-diacetate residue were performed. Cytotoxicity of the fluorescent cisplatin analogue in A2780 and A2780cis cells was determined using an MTT-based assay. The significance of differences was analysed using Student's t test or Mann-Whitney test as appropriate, p values of < 0.05 were considered significant. In the sensitive cells, both transporters are mainly localised in the trans-Golgi network, whereas they are sequestrated in more peripherally located vesicles in the resistant cells. Altered localisation of ATP7A and ATP7B in A2780cis cells is likely to be a consequence of major abnormalities in intracellular protein trafficking related to a reduced lysosomal compartment in this cell line. Changes in sub cellular localisation of ATP7A and ATP7B may facilitate sequestration of cisplatin in the vesicular structures of A2780cis cells, which may prevent drug binding to genomic DNA and thereby contribute to cisplatin resistance. Our results indicate that alterations in sub cellular localisation of transport proteins may contribute to cisplatin resistance in vitro. Investigation of intracellular protein localisation in primary tumour cell cultures and tumour tissues may help to develop markers of clinically relevant cisplatin resistance. Detection of resistant tumours in patients may in turn enable individualization of the chemotherapy in the early stage of treatment.
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影响因子：
64.5
作者：
Katzmann, DJ;Babst, M;Emr, SD
通讯作者：
Emr, SD
影响因子：
3
作者：
Hall, MD;Dillon, CT;Hambley, TW
通讯作者：
Hambley, TW
影响因子：
11.4
作者：
MOLLOY, SS;THOMAS, L;THOMAS, G
通讯作者：
THOMAS, G
影响因子：
3.5
作者：
REAVES, B;BANTING, G
通讯作者：
BANTING, G
人乳腺肿瘤细胞中的酸化不足及其对化学疗法的影响。
影响因子：
15.3
作者：
Altan, N;Chen, Y;Schindler, M;Simon, S M
通讯作者：
Simon, S M