顺铂诱导耐药蛋白BCRP与EHD2表达上调并在细胞表面的富集

李盼; 潘利; 付欣; 杨少斌; 冯婧文; 胡明秋; 应国光

doi:10.3969/j.issn.1000-8179.20131427

顺铂诱导耐药蛋白BCRP与EHD2表达上调并在细胞表面的富集

Cisplatin-induced up-regulation and enrichment of BCRP and EHD2 on cell surface

摘要

摘要:
目的建立顺铂耐受的人肺腺癌A549/DDP细胞株，研究EHD2（EH domain-containing protein-2）与肿瘤细胞药物耐受的潜在关系。
方法以DDP为诱导剂，采用低浓度逐步增量诱导方法建立人肺癌耐药细胞系A549/DDP，采用MTT法测定药物敏感性，免疫印迹法检测耐药蛋白BCRP及EHD2蛋白的表达水平，免疫荧光方法检测BCRP及EHD2的细胞定位。
结果成功建立DDP耐药细胞系A549/DDP，对DDP的耐药指数为7.6；免疫印迹结果显示BCRP在耐药细胞系A549/DDP中的表达明显高于其亲本细胞A549，EHD2蛋白在耐药细胞系的表达略有增高；免疫荧光成像显示EHD2蛋白在细胞膜表面富集，并与BCRP耐药蛋白共定位。
结论 BCRP和EHD2在获得性耐药细胞中富集，并于细胞膜表面共定位，提示内吞蛋白EHD2可能调控耐药蛋白BCRP的膜稳定性，从而介导肿瘤细胞耐药。

Abstract:
Objective To establish the cisplatin-resistant human lung adenocarcinoma cell line A549/(DDP) cisplatin and to study the relationship between EHD2 and drug resistance.
Methods DDP-resistant human lung cancer cell line A549/DDP was established by gradual and stepwise dose enhancement. MTT was used to measure drug sensitivity. Western blot and immunofluorescence were used to evaluate expression and subcellular localization of EHD2 and breast cancer resistance protein (BCRP).
Results The DDP-resistant cell line A549/DDP was established, with a resistance index of 7.6. EHD2 and BCRP expressions both increased and were enriched on the cell surface membrane.
Conclusion Both EHD2 and BCRP expressions were enriched on the resistant cell surface membrane, suggesting that EHD2 endocytic protein stabilizes BCRP and is involved in drug resistance.

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