1. Characterization of ABC Transporters in EpiAirway™, a Cellular Model of Normal Human Bronchial Epithelium.
- Author
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Rotoli BM, Barilli A, Visigalli R, Ferrari F, Frati C, Lagrasta CA, Lascia MD, Riccardi B, Puccini P, and Dall'Asta V
- Subjects
- ATP Binding Cassette Transporter, Subfamily B genetics, ATP Binding Cassette Transporter, Subfamily B metabolism, ATP Binding Cassette Transporter, Subfamily G, Member 2 genetics, ATP Binding Cassette Transporter, Subfamily G, Member 2 metabolism, Blotting, Western, Cell Line, Tumor, Humans, Immunohistochemistry, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Respiratory Mucosa metabolism, Reverse Transcriptase Polymerase Chain Reaction, Bronchi metabolism, Epithelium metabolism
- Abstract
The ATP-binding cassette (ABC) transporters P-glycoprotein (MDR1/ ABCB1 ), multidrug resistance-associated protein 1 (MRP1/ ABCC1 ), and breast cancer resistance protein (BCRP/ ABCG2 ) 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.
- Published
- 2020
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