Rapid detection of ABC transporter interaction: potential utility in pharmacology.
Rapid detection of ABC transporter interaction: potential utility in pharmacology.
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快速检测ABC转运蛋白相互作用：药理学中的潜在效用。
DOI：
10.1016/j.vascn.2010.11.003
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发表时间：
2011-05
影响因子：
1.9
通讯作者：
Bates SE
中科院分区：
文献类型：
--
作者：
Robey RW;Lin B;Qiu J;Chan LL;Bates SE
已知ATP结合盒（ABC）转运蛋白P-糖蛋白（Pgp/ABCB 1）、多药耐药相关蛋白1（MRP 1/ABCC 1）和乳腺癌耐药蛋白（BCRP/ABCG 2）可转运多种结构多样的化合物。它们在血脑、母胎和血睾丸屏障的高水平表达以及它们在口服吸收中的预期作用表明ABC转运蛋白发挥重要的药理学作用。我们已经开发了一种方法来表征ABC转运蛋白的功能和抑制，使用具有荧光检测能力的自动化细胞计数器。使用表达P-gp、MRP 1或ABCG 2的稳定转染的HEK 293细胞进行试验，并在存在或不存在已知抑制剂的情况下检查荧光底物的转运，并与流式细胞仪获得的结果进行比较。然后计算在不存在抑制剂的情况下与荧光底物孵育的细胞相对于在存在抑制剂的情况下的细胞内荧光的倍数增加。对于表达MRP 1或ABCG 2的细胞，使用细胞计数器或流式细胞仪获得的倍数增加值相当;与细胞计数器相比，在流式细胞仪上读取表达P-gp的细胞时，观察到的倍数增加值略高。所描述的测定提供了一种廉价的检测方法，以帮助开发新的ABC转运蛋白抑制剂或表征潜在的药物-药物相互作用。
The ATP-binding cassette (ABC) transporters P-glycoprotein (Pgp/ABCB1), multidrug resistance-associated protein 1 (MRP1/ABCC1), and breast cancer resistance protein (BCRP/ABCG2) are known to transport a wide range of structurally diverse compounds. Their high level of expression at the blood-brain, maternal-fetal, and blood-testis barriers as well as their purported roles in oral absorption suggests that ABC transporters play important pharmacologic roles. We have developed a method to characterize the function and inhibition of ABC transporters using an automated cell counter with fluorescence detection capability. The assay was performed using stably-transfected HEK293 cells expressing P-gp, MRP1, or ABCG2 and examining transport of fluorescent substrates in the presence or absence of known inhibitors and compared to results obtained with a flow cytometer. Fold-increase in intracellular fluorescence was then calculated for cells incubated with fluorescent substrate in the absence of inhibitor versus in the presence of inhibitor. Fold-increase values obtained either with the cell counter or flow cytometer were comparable for cells expressing either MRP1 or ABCG2; slightly higher fold-increase values were observed when cells expressing P-gp were read on a flow cytometer compared to the cell counter. The assay described provides an inexpensive detection method to aid in the development of novel ABC transporter inhibitors or to characterize potential drug-drug interactions.
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影响因子：
11.5
作者：
Lagas, Jurjen S.;van Waterschoot, Robert A. B.;Schinkel, Alfred H.
通讯作者：
Schinkel, Alfred H.
影响因子：
2.7
作者：
Dogan, AL;Legrand, O;Marie, JP
通讯作者：
Marie, JP
影响因子：
11.5
作者：
Kuppens, Isa E. L. M.;Witteveen, Els O.;Schellens, Jan H. M.
通讯作者：
Schellens, Jan H. M.
DOI：
10.1073/pnas.96.7.3900
发表时间：
1999-03-30
影响因子：
11.1
作者：
Rao, VV;Dahlheimer, JL;Piwnica-Worms, D
通讯作者：
Piwnica-Worms, D
影响因子：
5.7
作者：
Marchetti, Serena;de Vries, Nienke A.;Schellens, Jan H. M.
通讯作者：
Schellens, Jan H. M.