ABCG2/BCRP expression modulates D-luciferin-based bioluminescence imaging
ABCG2/BCRP expression modulates D-luciferin-based bioluminescence imaging
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DOI：
10.1158/0008-5472.can-07-0944
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发表时间：
2007-10-01
期刊：
影响因子：
11.2
通讯作者：
Pomper, Martin G.
中科院分区：
文献类型：
--
作者：
Zhang, Yimao;Bressler, Joseph P.;Pomper, Martin G.
生物发光成像（BLI）对于发育过程中的转基因表达的研究以及在许多体内疾病（如癌症）模型中对于体外高通量药物筛选变得不可或缺。由于D-荧光素酶与萤火虫荧光素酶（fLuc）的反应产生足够长波长的光子以允许在完整动物中成像，因此使用该底物和酶对已成为用于在体内进行BLI的选择方法。我们现在表明，表达的ATP结合盒（ABC）家族转运蛋白ABCG 2/BCRP影响BLI信号输出的底物D-胰蛋白酶。体外研究表明，D-精氨酸是ABCG 2/BCRP的底物，但不是MDR 1 P-糖蛋白（ABCB 1/Pgp）、多药耐药蛋白1（MRP 1/ABCC 1）或多药耐药蛋白2（MRP 2/ABCC 2）的底物。细胞内的D-荧光素摄取被ABC转运蛋白抑制剂调节，包括有效和选择性ABCG 2/BCRP抑制剂烟曲霉素C。经工程改造以表达转基因ABCG 2/BCRP的异种移植物以及源自天然表达ABCG 2/BCRP的人前列腺癌细胞系22 Rv 1的异种移植物的图像显示，目标区域内的ABCG 2/ BCRP表达和功能显著影响体内D-精氨酸依赖性生物发光输出。这些发现强调了在基于D-麦考蛋白的BLI期间考虑ABCG 2/BCRP效应的必要性，并提出了用于鉴定新的ABCG 2/ BCRP抑制剂的新型高通量方法。
Bioluminescence imaging (BLI) is becoming indispensable to the study of transgene expression during development and, in many in vivo models of disease such as cancer, for high throughput drug screening in vitro. Because reaction of D-luciferin with firefly luciferase (fLuc) produces photons of sufficiently long wavelength to permit imaging in intact animals, use of this substrate and enzyme pair has become the method of choice for performing BLI in vivo. We now show that expression of the ATP-binding cassette (ABC) family transporter ABCG2/BCRP affects BLI signal output from the substrate D-luciferin. In vitro studies show that D-luciferin is a substrate for ABCG2/BCRP but not for the MDR1 P-glycoprotein (ABCB1/Pgp), multidrug resistance protein 1 (MRP1/ABCC1), or multidrug; resistance protein 2 (MRP2/ABCC2). D-Luciferin uptake within cells is shown to be modulated by ABC transporter inhibitors, including the potent and selective ABCG2/BCRP inhibitor fumitremorgin C. Images of xenografts engineered to express transgenic ABCG2/BCRP, as well as xenografts derived from the human prostate cancer cell line 22Rv1 that naturally express ABCG2/BCRP, show that ABCG2/ BCRP expression and function within regions of interest substantially influence D-luciferin-dependent bioluminescent output in vivo. These findings highlight the need to consider ABCG2/BCRP effects during D-luciferin-based BLI and suggest novel high throughput methods for identifying new ABCG2/ BCRP inhibitors.