Development of analysis for multidrug resistance protein 1 (MRP1) modulator

多药耐药蛋白 1 (MRP1) 调节剂分析的进展

基本信息

批准号：
23791162
负责人：
OZEKI Michio
金额：
$ 2.58万
依托单位：
Gifu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2011
资助国家：
日本
起止时间：
2011 至 2012
项目状态：
已结题

项目摘要

Our purpose of research is to clarify the mechanism of anticancer drug resistanceby tumor cells and developing effective resistance modulators. Multidrug resistance can be mediated by overexpression of the multidrug resistance protein 1(MRP1). MRP function as transmembrane efflux pumps, which decrease intracellular drug accumulation, thereby conferring multidrug resistance. One of the ways to overcome MRP1 mediated multidrug resistance is to use an inhibitor to block the function of MRP1. This is called MRP1 modulator. To date, several MRP1 modulator have entered study and clinical trials. Recently, leukotriene receptor antagonist (LTRA) is thought to be one of the MRP1 modulator. We studied effect to overcome drug resistance by LTRA and have developed original candidate medications.In our study duration, we got results as below; The resistance cancer cells was established by two methods. The cells were selected from Jurkat (human leukemia cell line) by chronic exposure to doxorubicin over 2 months and transfection of MRP1cDNA. In the resistant cells, the overexpression of MRP1 resulted from an increased MRPmRNA level transcribed from amplified MRP gene. The dose-response effects of LTRA in the presence or absence of doxorubicin were examined in both drug-sensitive Jurkat and MRP-overexpressing resistant cells. LTRA reversed Jurkat resistance. The fluorescent accumulation analysis revealed a significant increase of fluorescence in resistant Jurkat pre-incubated LTRA. We demonstrated that LTRA modulate MRP1 scientifically by the measurement of intracellular glutathione, ATPase assay, analysis of cell-cycle pathway. The results were important findings for oncologic research. We have written the report now.

我们的研究目的是阐明肿瘤细胞抗癌药物耐药的机制并开发有效的耐药调节剂。多药耐药性可以通过多药耐药蛋白 1 (MRP1) 的过度表达来介导。 MRP 作为跨膜外排泵发挥作用，减少细胞内药物积累，从而赋予多药耐药性。克服MRP1介导的多药耐药性的方法之一是使用抑制剂来阻断MRP1的功能。这称为 MRP1 调制器。迄今为止，几种MRP1调节剂已进入研究和临床试验。最近，白三烯受体拮抗剂(LTRA)被认为是MRP1调节剂之一。我们研究了LTRA克服耐药性的效果，并开发了原创候选药物。在我们的研究期间，我们得到了以下结果；通过两种方法建立耐药癌细胞。通过长期暴露于阿霉素超过 2 个月并转染 MRP1cDNA，从 Jurkat（人类白血病细胞系）中选择细胞。在耐药细胞中，MRP1 的过度表达是由于扩增的 MRP 基因转录的 MRPmRNA 水平增加所致。在药物敏感的 Jurkat 和 MRP 过表达的耐药细胞中检查了在存在或不存在阿霉素的情况下 LTRA 的剂量反应效应。 LTRA 逆转了 Jurkat 的抵抗。荧光累积分析显示，抗性 Jurkat 预孵育 LTRA 中的荧光显着增加。我们通过细胞内谷胱甘肽的测量、ATP酶测定、细胞周期通路分析证明LTRA科学地调节MRP1。这些结果是肿瘤学研究的重要发现。我们现在已经写好了报告。