Modulation of function of three ABC drug transporters, P-glycoprotein (ABCB1), mitoxantrone resistance protein (ABCG2) and multidrug resistance protein 1 (ABCC1) by tetrahydrocurcumin, a major metabolite of curcumin
Modulation of function of three ABC drug transporters, P-glycoprotein (ABCB1), mitoxantrone resistance protein (ABCG2) and multidrug resistance protein 1 (ABCC1) by tetrahydrocurcumin, a major metabolite of curcumin
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DOI：
10.1007/s11010-006-9302-8
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发表时间：
2007-02-01
影响因子：
4.3
通讯作者：
Ambudkar, Suresh V.
中科院分区：
文献类型：
--
作者：
Limtrakul, Pornngarm;Chearwae, Wanida;Ambudkar, Suresh V.
许多研究的目的是开发有效的耐药调节剂，以克服人类癌症的多药耐药（MDR）。有效的耐多药调节剂正在临床试验中进行研究。许多目前的研究都集中在膳食草药上，因为这些草药已经使用了几个世纪，没有产生任何有害的副作用。在本研究中，研究了四氢姜黄素（THC）对三种ABC药物转运蛋白，p -糖蛋白（P-gp或ABCB1），米托蒽醌耐药蛋白（MXR或ABCG2）和多药耐药蛋白1 （MRP1或ABCC1）的影响，以评估姜黄素（THC）的最终代谢产物形式是否能够调节肿瘤细胞的MDR。两种不同类型的细胞系分别用于P-gp研究，人宫颈癌KB-3-1（野生型）和KB-V-1和人乳腺癌MCF-7（野生型）和MCF-7 MDR，而pcDNA3.1和pcDNA3.1-MRP1分别转染HEK 293和MXR过表达MCF7AdrVp3000或MCF7FL1000及其亲本MCF-7用于MRP1和MXR研究。我们首次报道了THC能够抑制P-gp、MXR和MRP1的功能。流式细胞术检测结果表明，四氢大麻酚能够抑制P-gp的功能，从而显著增加KB-V-1细胞中罗丹明和钙黄蛋白AM的积累。四氢大麻酚对MCF-7和MCF-7MDR中[H-3]-长春花碱积累和外排的影响证实了这一结果。四氢大麻酚显著增加MCF-7 MDR中[H-3]-长春花碱的积累并抑制其外排，且呈浓度依赖性。这种效应在野生型MCF-7细胞系中未发现。atp酶测定和P-gp与转运底物的光亲和标记清楚地表明THC与P-gp分子的相互作用。四氢大麻酚刺激P-gp atp酶活性，并以浓度依赖性的方式抑制[I-125]-碘arylazidoprazosin （IAAP）进入P-gp。THC也抑制了[I-125]-IAAP与MXR的结合，提示THC与转运体的药物结合位点相互作用。四氢大麻酚剂量依赖性地抑制MXR表达细胞（MCF7AdrVp3000和MCF7FL1000）的米托蒽醌和磷素A的外排。与MRP1类似，四氢大麻酚能有效抑制荧光底物钙黄蛋白AM的外排，从而增加MRP1- hek 293细胞中钙黄蛋白的积累，而不是其亲本pcDNA3.1-HEK 293细胞。MTT法测定四氢大麻酚对P-gp、MRP1和MXR的MDR逆转性能。四氢大麻酚可显著提高长春花碱、米托蒽醌和依托泊苷对耐药细胞KB-V-1、MCF7AdrVp3000和MRP1-HEK 293的敏感性。在各自的药物敏感亲本细胞系中没有发现这种效应。综上所述，本研究清楚地表明，四氢大麻酚抑制P-gp、MXR和MRP1的外排功能，能够延长姜黄素在体内的MDR逆转活性。
Many studies have been performed with the aim of developing effective resistance modulators to overcome the multidrug resistance (MDR) of human cancers. Potent MDR modulators are being investigated in clinical trials. Many current studies are focused on dietary herbs due to the fact that these have been used for centuries without producing any harmful side effects. In this study, the effect of tetrahydrocurcumin (THC) on three ABC drug transporter proteins, P-glycoprotein (P-gp or ABCB1), mitoxantrone resistance protein (MXR or ABCG2) and multidrug resistance protein 1 (MRP1 or ABCC1) was investigated, to assess whether an ultimate metabolite form of curcuminoids (THC) is able to modulate MDR in cancer cells. Two different types of cell lines were used for P-gp study, human cervical carcinoma KB-3-1 (wild type) and KB-V-1 and human breast cancer MCF-7 (wild type) and MCF-7 MDR, whereas, pcDNA3.1 and pcDNA3.1-MRP1 transfected HEK 293 and MXR overexpressing MCF7AdrVp3000 or MCF7FL1000 and its parental MCF-7 were used for MRP1 and MXR study, respectively. We report here for the first time that THC is able to inhibit the function of P-gp, MXR and MRP1. The results of flow cytometry assay indicated that THC is able to inhibit the function of P-gp and thereby significantly increase the accumulation of rhodamine and calcein AM in KB-V-1 cells. The result was confirmed by the effect of THC on [H-3]-vinblastine accumulation and efflux in MCF-7 and MCF-7MDR. THC significantly increased the accumulation and inhibited the efflux of [H-3]-vinblastine in MCF-7 MDR in a concentration-dependent manner. This effect was not found in wild type MCF-7 cell line. The interaction of THC with the P-gp molecule was clearly indicated by ATPase assay and photoaffinity labeling of P-gp with transport substrate. THC stimulated P-gp ATPase activity and inhibited the incorporation of [I-125]-iodoarylazidoprazosin (IAAP) into P-gp in a concentration-dependent manner. The binding of [I-125]-IAAP to MXR was also inhibited by THC suggesting that THC interacted with drug binding site of the transporter. THC dose dependently inhibited the efflux of mitoxantrone and pheophorbide A from MXR expressing cells (MCF7AdrVp3000 and MCF7FL1000). Similarly with MRP1, the efflux of a fluorescent substrate calcein AM was inhibited effectively by THC thereby the accumulation of calcein was increased in MRP1-HEK 293 and not its parental pcDNA3.1-HEK 293 cells. The MDR reversing properties of THC on P-gp, MRP1, and MXR were determined by MTT assay. THC significantly increased the sensitivity of vinblastine, mitoxantrone and etoposide in drug resistance KB-V-1, MCF7AdrVp3000 and MRP1-HEK 293 cells, respectively. This effect was not found in respective drug sensitive parental cell lines. Taken together, this study clearly showed that THC inhibits the efflux function of P-gp, MXR and MRP1 and it is able to extend the MDR reversing activity of curcuminoids in vivo.