The exposure of cancer cells to hyperthermia, iron oxide nanoparticles, and mitomycin C influences membrane multidrug resistance protein expression levels.
The exposure of cancer cells to hyperthermia, iron oxide nanoparticles, and mitomycin C influences membrane multidrug resistance protein expression levels.
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DOI：
10.2147/ijn.s37465
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发表时间：
2013
影响因子：
8
通讯作者：
Hilger I
中科院分区：
文献类型：
--
作者：
Franke K;Kettering M;Lange K;Kaiser WA;Hilger I
多药耐药相关蛋白（MRP）在癌细胞中的存在是已知的负责在目前的肿瘤治疗中的许多治疗失败。在这里，我们表明，不同的效应器，如热疗，氧化铁纳米粒子和化疗药物的组合影响MRP 1和3在腺癌细胞系中的表达。在存在或不存在热疗（43°C，15至120分钟）的情况下，用磁性纳米颗粒（MNP; 1.5至150 μg Fe/cm 2）或丝裂霉素C（高达1.5 μg/cm 2，24小时）处理BT-474细胞。此外，细胞也依次暴露于这些效应物（MNP、高热和丝裂霉素C）。细胞收获后，提取mRNA并通过逆转录聚合酶链反应进行分析。此外，膜蛋白的分离和分析，通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹。当细胞暴露于单独的效应物或其组合时，未观察到对MRP 1和3 mRNA表达的影响。相反，膜蛋白表达的影响，以选择性的方式。与MRP 1相比，对MRP 3表达的影响不太明显。丝裂霉素C治疗降低MRP的表达在高浓度和高温加强这些影响。与此相反，MNP的存在只增加MRP 1和3的表达，和高温逆转这些影响。当结合热疗，磁性纳米粒子，丝裂霉素C，没有进一步抑制MRP表达的观察相比，各自的双重治疗方式。不同的MRP 1和3表达水平与从头mRNA表达无关，而是与由于活性氧产生导致的MRP 1和3向细胞膜的改变的易位有关，并且与细胞内MRP储存库的移位、膜流动性的变化等在蛋白质水平上有关。我们的研究结果可用于开发新的治疗策略，通过抑制主动从靶细胞输出药物的机制，从而改善肿瘤学的治疗效果。
The presence of multidrug resistance-associated protein (MRP) in cancer cells is known to be responsible for many therapeutic failures in current oncological treatments. Here, we show that the combination of different effectors like hyperthermia, iron oxide nanoparticles, and chemotherapeutics influences expression of MRP 1 and 3 in an adenocarcinoma cell line. BT-474 cells were treated with magnetic nanoparticles (MNP; 1.5 to 150 μg Fe/cm2) or mitomycin C (up to 1.5 μg/cm2, 24 hours) in the presence or absence of hyperthermia (43°C, 15 to 120 minutes). Moreover, cells were also sequentially exposed to these effectors (MNP, hyperthermia, and mitomycin C). After cell harvesting, mRNA was extracted and analyzed via reverse transcription polymerase chain reaction. Additionally, membrane protein was isolated and analyzed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. When cells were exposed to the effectors alone or to combinations thereof, no effects on MRP 1 and 3 mRNA expression were observed. In contrast, membrane protein expression was influenced in a selective manner. The effects on MRP 3 expression were less pronounced compared with MRP 1. Treatment with mitomycin C decreased MRP expression at high concentrations and hyperthermia intensified these effects. In contrast, the presence of MNP only increased MRP 1 and 3 expression, and hyperthermia reversed these effects. When combining hyperthermia, magnetic nanoparticles, and mitomycin C, no further suppression of MRP expression was observed in comparison with the respective dual treatment modalities. The different MRP 1 and 3 expression levels are not associated with de novo mRNA expression, but rather with an altered translocation of MRP 1 and 3 to the cell membrane as a result of reactive oxygen species production, and with shifting of intracellular MRP storage pools, changes in membrane fluidity, etc, at the protein level. Our results could be used to develop new treatment strategies by repressing mechanisms that actively export drugs from the target cell, thereby improving the therapeutic outcome in oncology.