Oxidative Stress Promotes Doxorubicin-Induced Pgp and BCRP Expression in Colon Cancer Cells Under Hypoxic Conditions
Oxidative Stress Promotes Doxorubicin-Induced Pgp and BCRP Expression in Colon Cancer Cells Under Hypoxic Conditions
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DOI：
10.1002/jcb.25890
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Garzon, Ruth
中科院分区：
文献类型：
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作者：
Pinzon-Daza, Martha L.;Cuellar-Saenz, Yenith;Garzon, Ruth
P-糖蛋白（Pgp）和乳腺癌耐药蛋白（BCRP）是ATP结合盒（ABC）转运蛋白，在不同的耐药癌细胞系中过表达。在这项研究中，我们调查是否阿霉素促进Pgp和/或BCRP的表达，诱导耐药的结肠癌细胞在缺氧条件下。我们通过ELISA分析了HIF-1 α活性，通过qRT-PCR分析了Pgp和BCRP表达，并通过共聚焦显微镜分析了HT-29 WT和HT-29多柔比星耐药结肠癌细胞（HT-29 DxR）中多柔比星摄取与ABC转运蛋白表达之间的关系。这些细胞用阿霉素和/或CoCl 2（化学缺氧）和活性氧诱导剂处理。我们发现，化学诱导的缺氧和阿霉素的组合促进Pgp mRNA的表达在24小时内在HT-29 WT和HT-29 DxR细胞。阿霉素和CoCl 2单独或联合诱导Pgp和BCRP表达，如上述两种细胞系中每种细胞系的共聚焦显微镜所示。因此，我们推测，Pgp和BCRP的表达可能是由于在缺氧条件下阿霉素诱导的ROS产生和HIF-1a活性的联合作用产生的协同效应。然而，HIF-1a活性中断通过管理的APE-1抑制剂E3330，下调Pgp表达和增加阿霉素交付HT-29细胞，在那里它作为Pgp的底物，表明Pgp表达和阿霉素积累之间存在间接关系。因此，我们得出结论，Pgp和BCRP表达可通过阿霉素和缺氧之间的相互作用调节，促进HT-29 WT和HT-29 DxR细胞的耐药性，并且该过程可能是ROS依赖性的。(C)2017 Wiley Periodicals，Inc.
P-glycoprotein (Pgp) and breast cancer resistance protein (BCRP) are ATP binding cassette (ABC) transporters that are overexpressed in different drug-resistant cancer cell lines. In this study, we investigated whether doxorubicin promotes Pgp and/or BCRP expression to induce drug resistance in colon cancer cells under hypoxic conditions. We analyzed HIF-1 alpha activity via ELISA, Pgp, and BCRP expression by qRT-PCR and the relationship between doxorubicin uptake and ABC transporter expression via confocal microscopy in HT-29WT and HT-29 doxorubicin-resistant colon cancer cells (HT-29DxR). These cells were treated with doxorubicin and/or CoCl2 (chemical hypoxia), and reactive oxygen species inductors. We found that the combination of chemically induced hypoxia and doxorubicin promoted Pgp mRNA expression within 24 h in HT-29WT and HT-29DxR cells. Both doxorubicin and CoCl2 alone or in combination induced Pgp and BCRP expression, as demonstrated via confocal microscopy in each of the above two cell lines. Thus, we surmised that Pgp and BCRP expression may result from synergistic effects exerted by the combination of doxorubicin-induced ROS production and HIF-1a activity under hypoxic conditions. However, HIF-1a activity disruption via the administration of E3330, an APE-1 inhibitor, downregulated Pgp expression and increased doxorubicin delivery to HT-29 cells, where it served as a substrate for Pgp, indicating the existence of an indirect relationship between Pgp expression and doxorubicin accumulation. Thus, we concluded that Pgp and BCRP expression can be regulated via cross-talk between doxorubicin and hypoxia, promoting drug resistance in HT-29 WT, and HT-29DxR cells and that this process may be ROS dependent. (C) 2017 Wiley Periodicals, Inc.