In vitro study of P-glycoprotein induction as an antidotal pathway to prevent cytotoxicity in Caco-2 cells

• Type: Article in International Scientific Journal
• Date: 2011

Authors

Renata Silva (Author) Other; Helena Carmo (Author) FFUP; Ricardo Dinis Oliveira (Author) FMUP; Anabela Cordeiro da Silva (Author) FFUP; Sofia Costa Lima (Author) Other; Felix Carvalho (Author) FFUP; Maria de Lourdes Bastos (Author) FFUP; Fernando Remiao (Author) FFUP

Journal

Title: Archives of Toxicology

Vol. 85

Pages: 315-326

ISSN: 0340-5761

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 44 Citations

Scopus - 50 Citations

Scientific classification

FOS: Medical and Health sciences > Basic medicine

Other information

Authenticus ID: P-002-STX

DOI: 10.1007/s00204-010-0587-8

Abstract (EN): The Caco-2 cell line is a reliable in vitro model for predicting drug intestinal absorption and P-glycoprotein (P-gp)-mediated excretion in humans. Recent in vivo studies suggested the induction of P-gp as a cellular protection tool against paraquat poisoning, through the increase in its pulmonary and intestinal excretion. Thus, the aim of the present work was to evaluate P-gp expression and activity in Caco-2 cells exposed to doxorubicin (a known P-gp inducer) and to correlate these changes with paraquat toxic effects. Cytotoxicity of doxorubicin (0-100 mu M) and paraquat (0-1,000 mu M) was evaluated for a maximum period of 96 h. In doxorubicin-exposed cells, P-gp expression and transport activity were evaluated by flow cytometry, using a fluorescein isothiocyanate-conjugated antibody and the P-gp fluorescent subtract rhodamine 123, respectively. A significant increase in P-gp expression was observed as soon as 6 h after exposure to 5 mu M doxorubicin. P-gp activity also increased after 6 h, but only at higher doxorubicin concentrations (over 50 mu M). Paraquat (0-5,000 mu M) cytotoxicity was then evaluated with or without previous exposure of the cells to doxorubicin (5-100 mu M, a concentration range causing both an increase in P-gp expression and activity). Under P-gp induction, a significant reduction in paraquat cytotoxicity was observed. Furthermore, when these cells were incubated with a specific P-gp inhibitor (UIC2 antibody) the doxorubicin protective effects were blocked, confirming the involvement of P-gp in the reduction in paraquat cytotoxicity. In conclusion, the human Caco-2 cell line model can be used for the study of P-gp induction as an antidotal pathway against substrates of this transporter system.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 12