Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, Via Volturno, 39, 43125 Parma, Italy.
Pathology Unit, Department of Medicine and Surgery, University of Parma, Via Gramsci, 14, 43126 Parma, Italy.
Int J Mol Sci. 2020 Apr 30;21(9):3190. doi: 10.3390/ijms21093190.
The ATP-binding cassette (ABC) transporters P-glycoprotein (MDR1/), multidrug resistance-associated protein 1 (MRP1/), and breast cancer resistance protein (BCRP/) play a crucial role in the translocation of a broad range of drugs; data about their expression and activity in lung tissue are controversial. Here, we address their expression, localization and function in EpiAirway™, a three-dimensional (3D)-model of human airways; Calu-3 cells, a representative in vitro model of bronchial epithelium, are used for comparison. Transporter expression has been evaluated with RT-qPCR and Western blot, the localization with immunocytochemistry, and the activity by measuring the apical-to-basolateral and basolateral-to-apical fluxes of specific substrates in the presence of inhibitors. EpiAirway™ and Calu-3 cells express high levels of MRP1 on the basolateral membrane, while they profoundly differ in terms of BCRP and MDR1: BCRP is detected in EpiAirway™, but not in Calu-3 cells, while MDR1 is expressed and functional only in fully-differentiated Calu-3; in EpiAirway™, MDR1 expression and activity are undetectable, consistently with the absence of the protein in specimens from human healthy bronchi. In summary, EpiAirway™ appears to be a promising tool to study the mechanisms of drug delivery in the bronchial epithelium and to clarify the role of ABC transporters in the modulation of the bioavailability of administered drugs.
三磷酸腺苷结合盒（ABC）转运蛋白 P-糖蛋白（MDR1/）、多药耐药相关蛋白 1（MRP1/）和乳腺癌耐药蛋白（BCRP/）在广泛的药物转运中发挥着关键作用；关于它们在肺组织中的表达和活性的数据存在争议。在这里，我们研究了它们在三维（3D）人呼吸道模型 EpiAirway™中的表达、定位和功能；Calu-3 细胞是支气管上皮的代表性体外模型，用于比较。使用 RT-qPCR 和 Western blot 评估转运蛋白的表达，用免疫细胞化学评估定位，通过在存在抑制剂的情况下测量特定底物的顶端到基底和基底到顶端的通量来评估活性。EpiAirway™和 Calu-3 细胞在基底外侧膜上表达高水平的 MRP1，而在 BCRP 和 MDR1 方面则存在显著差异：BCRP 在 EpiAirway™中检测到，但在 Calu-3 细胞中未检测到，而 MDR1 仅在完全分化的 Calu-3 中表达和有功能；在 EpiAirway™中，MDR1 的表达和活性无法检测到，与来自健康人支气管的标本中不存在该蛋白一致。总之，EpiAirway™似乎是研究支气管上皮药物输送机制和阐明 ABC 转运蛋白在调节给药药物生物利用度中的作用的有前途的工具。
