Archives of Toxicology

, Volume 92, Issue 6, pp 2027–2042 | Cite as

Using the lentiviral vector system to stably express chicken P-gp and BCRP in MDCK cells for screening the substrates and studying the interplay of both transporters

  • Yujuan Zhang
  • Jinhu Huang
  • Yang Liu
  • Tingting Guo
  • Liping WangEmail author
Molecular Toxicology


Transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are known to influence the pharmacokinetics and toxicity of substrate drugs. However, no detailed information is as yet available about functional activity and substrate spectra of chicken P-gp and BCRP. In this study, BCRP single and BCRP/P-gp double-transfected MDCK cell lines (named MDCK-chAbcg2 and MDCK-chAbcg2/Abcb1, respectively) were generated using lentiviral vector system to develop reliable systems for screening the substrates for these two transporters and study the interplay between them. The constructed cell lines significantly expressed functional exogenous proteins and expression persisted for at least 50 generations with no decrease. Enrofloxacin, ciprofloxacin, tilmicosin, sulfadiazine, ampicillin and clindamycin were classified as the substrates of chicken P-gp according to the rules suggested by FDA, as their net efflux ratios were greater than two. Similarly, enrofloxacin, ciprofloxacin, tilmicosin, florfenicol, ampicillin and clindamycin were classified as the substrates of BCRP. Among these drugs, enrofloxacin, ciprofloxacin, tilmicosin, ampicillin, and clindamycin were the cosubstrates of P-gp and BCRP, however, chicken BCRP and P-gp exhibit different affinities to the shared substrates at different concentrations by blocking either one or both transport with specific inhibitors in the coexpression system. It was also found that ceftiofur, amoxicillin and doxycycline were not substrates of either chicken BCRP or the substrates of chicken P-gp. These constructed cell models provide useful systems for high-throughput screening of the potential substrates of chicken BCRP and P-gp as well as the drug–drug interaction mediated via chicken BCRP and P-gp.


Chicken BCRP Chicken P-gp MDCK Lentiviral vector system Drug–drug interaction 



This work was supported by the National Key Research and Development Program (2016YFD0501309), Postgraduate Research & Practice Innovation Program of Jiangsu Province and Qinlan Project of Jiangsu Province (2014).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

204_2018_2209_MOESM1_ESM.docx (386 kb)
Supplementary material 1 (DOCX 386 KB)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yujuan Zhang
    • 1
  • Jinhu Huang
    • 1
  • Yang Liu
    • 1
  • Tingting Guo
    • 1
  • Liping Wang
    • 1
    Email author
  1. 1.Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary MedicineNanjing Agricultural UniversityNanjingPeople’s Republic of China

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