ATP-Binding Cassette C Transporters in Human Pancreatic Carcinoma Cell Lines
ATP-Binding Cassette C Transporters in Human Pancreatic Carcinoma Cell Lines
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DOI：
10.1159/000178884
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发表时间：
2009-01-01
期刊：
影响因子：
3.6
通讯作者：
Lohr, J. Matthias
中科院分区：
文献类型：
--
作者：
Hagmann, Wolfgang;Jesnowski, Ralf;Lohr, J. Matthias
背景：胰腺癌的特点是对化疗具有高耐药性。这种化疗耐药性可以由多药耐药蛋白 (MRP)、乳腺癌耐药蛋白 (BCRP) 和 MDR1 P-糖蛋白介导。然而，单个 MRP 亚型对胰腺癌化疗耐药的贡献尚不清楚。我们研究了人胰腺癌细胞系与原代胰管细胞中 ATP 结合盒 (ABC) 转运蛋白的表达，并分析了 5-氟尿嘧啶耐药细胞中的 MRP 表达谱。方法：通过定量和定性 RT-PCR、免疫印迹以及细胞毒性测定来分析转运蛋白表达。结果：原代胰管细胞表达MRP1、MRP3、MRP4和MRP5，但不表达MRP2 mRNA。建立的癌细胞系表达MRP1、MRP4和MRP5，其中大多数还表达MRP2、MRP3、MRP7和BCRP，但没有一个含有可检测量的MRP6、MRP8或MRP9 mRNA。免疫印迹分析表明，所有细胞中都存在 MRP1、MRP4 和 MRP5 蛋白，但仅在其中一些细胞中存在 MRP3 和 BCRP 蛋白。与亲本 Capan-1 细胞相比，对 5-氟尿嘧啶获得性化学耐药的 Capan-1 细胞显示 MRP3、MRP4 和 MRP5 的 mRNA 和蛋白表达上调。此外，通过RNA干扰沉默MRP5导致亲代Capan-1细胞对5-氟尿嘧啶细胞毒性的敏感性增强。结论：MRP3、MRP4和MRP5在5-氟尿嘧啶耐药细胞中表达上调，MRP5有助于胰腺癌细胞对5-FU耐药。版权所有 (C) 2008 S. Karger AG，巴塞尔和 IAP
Background: Pancreatic cancer is characterized by high resistance to chemotherapy. Such chemoresistance can be mediated by multidrug resistance proteins (MRPs), breast cancer resistance protein (BCRP), and MDR1 P-glycoprotein. However, the contribution of individual MRP isoforms to chemoresistance in pancreatic carcinoma is unclear. We studied ATP-binding cassette (ABC) transporter expression in human pancreatic carcinoma cell lines as compared to primary pancreatic duct cells, and analyzed the MRP expression profile in 5-fluorouracil-resistant cells. Methods: Transporter expression was analyzed by quantitative and qualitative RT-PCR, by immunoblot, and chemoresistance by cytotoxicity assay. Results: Primary pancreatic duct cells expressed MRP1, MRP3, MRP4, and MRP5, but not MRP2 mRNA. The established carcinoma cell lines expressed MRP1, MRP4, and MRP5, most of them also MRP2, MRP3, MRP7, and BCRP, but none contained detectable amounts of MRP6, MRP8, or MRP9 mRNA. Immunoblot analyses demonstrated presence of MRP1, MRP4, and MRP5 protein in all, but MRP3 and BCRP protein only in some of these cells. Compared to parental Capan-1 cells, Capan-1 cells with acquired chemoresistance towards 5-fluorouracil showed an upregulated mRNA and protein expression of MRP3, MRP4, and MRP5. In addition, silencing of MRP5 by RNA interference resulted in enhanced sensitivity of parental Capan-1 cells towards 5-fluorouracil cytotoxicity. Conclusion: MRP3, MRP4, and MRP5 are upregulated in 5-fluorouracil-resistant cells, and MRP5 contributes to 5-FU resistance in pancreatic carcinoma cells. Copyright (C) 2008 S. Karger AG, Basel and IAP