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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 Wang
Molecular Toxicology
  • 78 Downloads

Abstract

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.

Keywords

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

Notes

Acknowledgements

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)

References

  1. Cui Y, König J, Keppler D (2001) Vectorial transport by double-transfected cells expressing the human uptake transporter SLC21A8 and the apical export pump ABCC2. Mol Pharmacol 60:934CrossRefPubMedGoogle Scholar
  2. Demoling LA, Bååth E, Greve G, Wouterse M, Schmitt H (2009) Effects of sulfamethoxazole on soil microbial communities after adding substrate. Soil Biol Biochem 41:840–848.  https://doi.org/10.1016/j.soilbio.2009.02.001 CrossRefGoogle Scholar
  3. Fung KL, Kapoor K, Pixley JN, Talbert DJ, Kwit ADT, Ambudkar SV, Gottesman MM (2015) Using the BacMam Baculovirus system to study expression and function of recombinant efflux drug transporters in polarized epithelial cell monolayers. Drug Metab Dispos 44:180–188.  https://doi.org/10.1124/dmd.115.066506 CrossRefPubMedGoogle Scholar
  4. Gartzke D, Fricker G (2014) Establishment of optimized MDCK cell lines for reliable efflux transport studies. J Pharm Sci 103:1298–1304.  https://doi.org/10.1002/jps.23901 CrossRefPubMedGoogle Scholar
  5. Giacomini KM, Huang SM (2013) Transporters in drug development and clinical pharmacology clinical. Pharmacol Ther 94:3–9.  https://doi.org/10.1038/clpt.2013.86 CrossRefGoogle Scholar
  6. Griffiths NM, Hirst BH, Simmons NL (1993) Active secretion of the fluoroquinolone ciprofloxacin by human intestinal epithelial Caco-2 cell layers. Br J Pharmacol 108:575–576CrossRefPubMedPubMedCentralGoogle Scholar
  7. Guo M, Bughio S, Sun Y, Zhang Y, Dong L, Dai X, Wang L (2013) Age-related P-glycoprotein expression in the intestine and affecting the pharmacokinetics of orally administered enrofloxacin in broilers. Plos One 8:e74150CrossRefPubMedPubMedCentralGoogle Scholar
  8. Guo T et al. (2016) Abcb1 in Pigs: Molecular cloning, tissues distribution, functional analysis, and its effect on pharmacokinetics of enrofloxacin. Sci Rep  https://doi.org/10.1038/srep32244 Google Scholar
  9. Hammesfahr U, Heuer H, Manzke B, Smalla K, Thiele-Bruhn S (2008) Impact of the antibiotic sulfadiazine and pig manure on the microbial community structure in agricultural soils. Soil Biol Biochem 40:1583–1591.  https://doi.org/10.1016/j.soilbio.2008.01.010 CrossRefGoogle Scholar
  10. Haslam IS, Wright JA, O’Reilly DA, Sherlock DJ, Coleman T, Simmons NL (2011) Intestinal ciprofloxacin efflux: the role of breast cancer resistance protein (ABCG2). Drug Metab Dispos 39:2321–2328.  https://doi.org/10.1124/dmd.111.038323 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Kataoka M, Yokoyama T, Masaoka Y, Sakuma S, Yamashita S (2011) Estimation of P-glycoprotein-mediated efflux in the oral absorption of P-gp substrate drugs from simultaneous analysis of drug dissolution and permeation. Eur J Pharm Sci 44:544–551.  https://doi.org/10.1016/j.ejps.2011.09.007 CrossRefPubMedGoogle Scholar
  12. Kathawala RJ, Gupta P, Ashby CR, Chen Z-S (2015) The modulation of ABC transporter-mediated multidrug resistance in cancer: A review of the past decade. Drug Resist Updates 18:1–17.  https://doi.org/10.1016/j.drup.2014.11.002 CrossRefGoogle Scholar
  13. Kux L (2012) Draft guidance for industry on drug interaction studies-study design, data analysis, Implications for Dosing, and Labeling Recommendations; Availability Federal RegisterGoogle Scholar
  14. Lentz KA, Polli JW, Wring SA, Humphreys JE, Polli JE (2000) Influence of passive permeability on apparent P-glycoprotein kinetics. Pharm Res 17:1456–1460CrossRefPubMedGoogle Scholar
  15. Li M, de Graaf IAM, Siissalo S, de Jager MH, van Dam A, Groothuis GMM (2016) The Consequence of drug–drug interactions influencing the interplay between P-glycoprotein and cytochrome P450 3a: an ex vivo study with rat precision-cut intestinal slices. Drug Metab Dispos 44:683–691.  https://doi.org/10.1124/dmd.115.068684 CrossRefPubMedGoogle Scholar
  16. Li Y, Revalde J, Paxton JW (2017) The effects of dietary and herbal phytochemicals on drug transporters. Adv Drug Deliv Rev 116:45–62.  https://doi.org/10.1016/j.addr.2016.09.004 CrossRefPubMedGoogle Scholar
  17. Lin H et al (2016) Effects of manure and mineral fertilization strategies on soil antibiotic resistance gene levels and microbial community in a paddy–upland rotation system. Environ Pollut 211:332–337.  https://doi.org/10.1016/j.envpol.2016.01.007 CrossRefPubMedGoogle Scholar
  18. Merino G (2005) The breast cancer resistance protein (BCRP/ABCG2) affects pharmacokinetics, hepatobiliary excretion, and milk secretion of the antibiotic nitrofurantoin. Mol Pharmacol 67:1758–1764.  https://doi.org/10.1124/mol.104.010439 CrossRefPubMedGoogle Scholar
  19. Miyamoto R, Nozawa T, Kimura M, Shiozuka K, Tabata K (2015) Development and validation of semiautomated 96-well transport assay using LLC-PK1 cells transfected with human P-glycoprotein for high-throughput screening. ASSAY Drug Dev Technol 13:79–87.  https://doi.org/10.1089/adt.2014.621 CrossRefPubMedGoogle Scholar
  20. Montanari F, Ecker GF (2015) Prediction of drug–ABC–transporter interaction—recent advances and future challenges. Adv Drug Deliv Rev 86:17–26.  https://doi.org/10.1016/j.addr.2015.03.001 CrossRefPubMedGoogle Scholar
  21. Muenster U, Grieshop B, Ickenroth K, Gnoth MJ (2008) Characterization of substrates and inhibitors for the in vitro assessment of Bcrp mediated drug-drug interactions. Pharm Res 25:2320–2326.  https://doi.org/10.1007/s11095-008-9632-1 CrossRefPubMedGoogle Scholar
  22. Niu J et al (2013) Lentivirus-mediated CD/TK fusion gene transfection neural stem cell therapy for C6 glioblastoma. Tumor Biol 34:3731–3741.  https://doi.org/10.1007/s13277-013-0957-y CrossRefGoogle Scholar
  23. Omasits U, Ahrens CH, Müller S, Wollscheid B (2014) Protter: interactive protein feature visualization and integration with experimental proteomic data. Bioinformatics 30:884–886.  https://doi.org/10.1093/bioinformatics/btt607 CrossRefPubMedGoogle Scholar
  24. Peachey LE et al (2017) P-glycoproteins play a role in ivermectin resistance in cyathostomins. Int J Parasitol Drugs Drug Resist 7:388–398.  https://doi.org/10.1016/j.ijpddr.2017.10.006 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Perez M, Otero JA, Barrera B, Prieto JG, Merino G, Alvarez AI (2013) Inhibition of ABCG2/BCRP transporter by soy isoflavones genistein and daidzein: effect on plasma and milk levels of danofloxacin in sheep. Vet J 196:203–208.  https://doi.org/10.1016/j.tvjl.2012.09.012 CrossRefPubMedGoogle Scholar
  26. Poller B, Wagenaar E, Tang SC, Schinkel AH (2011) Double-transduced MDCKII cells to study human P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) interplay in drug transport across the blood–brain barrier. Mol Pharm 8:571–582.  https://doi.org/10.1021/mp1003898 CrossRefPubMedGoogle Scholar
  27. Shukla S, Schwartz C, Kapoor K, Kouanda A, Ambudkar SV (2011) Use of Baculovirus BacMam vectors for expression of ABC drug transporters in mammalian cells. Drug Metab Dispos 40:304–312.  https://doi.org/10.1124/dmd.111.042721 CrossRefPubMedGoogle Scholar
  28. Sun Y, Guo T, Guo D, Guo L, Chen L, Zhang Y, Wang L (2016) Establishment and characterization of an MDCK cell line stably-transfected with chicken Abcb1 encoding P-glycoprotein. Res Vet Sci 106:37–44.  https://doi.org/10.1016/j.rvsc.2016.03.004 CrossRefPubMedGoogle Scholar
  29. Takada T, Suzuki H, Sugiyama Y (2005) Characterization of polarized expression of point- or deletion-mutated human BCRP/ABCG2 in LLC-PK1 cells. Pharm Res 22:458–464.  https://doi.org/10.1007/s11095-004-1884-9 CrossRefPubMedGoogle Scholar
  30. Takeuchi T et al (2006) Establishment and characterization of the transformants stably-expressing MDR1 derived from various animal species in LLC-PK1. Pharm Res 23:1460–1472.  https://doi.org/10.1007/s11095-006-0285-7 CrossRefPubMedGoogle Scholar
  31. Tang F, Horie K, Borchardt RT (2002) Are MDCK Cells transfected with the human MDR1 gene a good model of the human intestinal mucosa? Pharm Res 19:765–772.  https://doi.org/10.1023/a:1016140429238 CrossRefPubMedGoogle Scholar
  32. Tanizaki J et al (2010) Synergistic antitumor effect of S-1 and HER2-targeting agents in gastric cancer with HER2 amplification. Mol Cancer Ther 9:1198–1207.  https://doi.org/10.1158/1535-7163.Mct-10-0045 CrossRefPubMedGoogle Scholar
  33. Tian Y, Qian S, Jiang Y, Shen Q, Zheng J, Zhou H, Zeng S (2013) The interaction between human breast cancer resistance protein (BCRP) and five bisbenzylisoquinoline alkaloids. Int J Pharm 453:371–379.  https://doi.org/10.1016/j.ijpharm.2013.05.053 CrossRefPubMedGoogle Scholar
  34. Tseng WC, Haselton FR, Giorgio TD (1997) Transfection by cationic liposomes using simultaneous single cell measurements of plasmid delivery and transgene expression. J Biol Chem 272:25641–25647CrossRefPubMedGoogle Scholar
  35. Wassermann L, Halwachs S, Lindner S, Honscha KU, Honscha W (2013) Determination of functional ABCG2 activity and assessment of drug–ABCG2 interactions in dairy animals using a novel MDCKII in vitro model. J Pharm Sci 102:772–784.  https://doi.org/10.1002/jps.23399 CrossRefPubMedGoogle Scholar
  36. Wepking C et al (2017) Exposure to dairy manure leads to greater antibiotic resistance and increased mass-specific respiration in soil microbial communities. Proc R Soc B Biol Sci  https://doi.org/10.1098/rspb.2016.2233 Google Scholar
  37. Wu Y et al (2015) Lentivirus mediated over expression of CGRP inhibited oxidative stress in Schwann cell line. Neurosci Lett 598:52–58.  https://doi.org/10.1016/j.neulet.2015.05.009 CrossRefPubMedGoogle Scholar
  38. Yang SH, Lee MG (2007) Dose-independent pharmacokinetics of clindamycin after intravenous and oral administration to rats: contribution of gastric first-pass effect to low bioavailability. Int J Pharm 332:17–23.  https://doi.org/10.1016/j.ijpharm.2006.11.019 CrossRefPubMedGoogle Scholar
  39. Ziv G, Shemtov M, Glickman A, Winkler M, Saran A (1995) Tilmicosin antibacterial activity and pharmacokinetics in cows. J Vet Pharmacol Ther 18:340–345CrossRefPubMedGoogle Scholar

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
  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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