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Physiological intestinal oxygen modulates the Caco-2 cell model and increases sensitivity to the phytocannabinoid cannabidiol

  • Tara Macpherson
  • Jane A. Armstrong
  • David N. Criddle
  • Karen L. Wright
Article

Abstract

The Caco-2 cell model is widely used as a model of colon cancer and small intestinal epithelium but, like most cell models, is cultured in atmospheric oxygen conditions (∼21%). This does not reflect the physiological oxygen range found in the colon. In this study, we investigated the effect of adapting the Caco-2 cell line to routine culturing in a physiological oxygen (5%) environment. Under these conditions, cells maintain a number of key characteristics of the Caco-2 model, such as increased formation of tight junctions and alkaline phosphatase expression over the differentiation period and maintenance of barrier function. However, these cells exhibit differential oxidative metabolism, proliferate less and become larger during differentiation. In addition, these cells were more sensitive to cannabidiol-induced antiproliferative actions through changes in cellular energetics: from a drop of oxygen consumption rate and loss of mitochondrial membrane integrity in cells treated under atmospheric conditions to an increase in reactive oxygen species in intact mitochondria in cells treated under low-oxygen conditions. Inclusion of an additional physiological parameter, sodium butyrate, into the medium revealed a cannabidiol-induced proliferative response at low doses. These effects could impact on its development as an anticancer therapeutic, but overall, the data supports the principle that culturing cells in microenvironments that more closely mimic the in vivo conditions is important for drug screening and mechanism of action studies.

Keywords

Colon Cannabinoid Normoxia Oxidative metabolism Butyrate 

Notes

Acknowledgments

T. Macpherson is partly funded by a Novartis Pharmaceuticals UK Ltd (Horsham, UK) studentship with matched funding from the Biotechnology and Biological Sciences Research Council (BBSRC, Swindon, UK). K. Wright is partly funded by a Peel Trust Lectureship from The Dowager Countess Eleanor Peel Trust. We would also like to thank Dr Dave Clancy for useful discussions on data interpretation and statistics and Dr Jane Andre for assistance with the confocal imaging (both from Faculty of Health and Medicine, Lancaster University).

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Copyright information

© The Society for In Vitro Biology 2014

Authors and Affiliations

  • Tara Macpherson
    • 1
  • Jane A. Armstrong
    • 2
  • David N. Criddle
    • 2
  • Karen L. Wright
    • 1
  1. 1.Faculty of Health and Medicine, Division of Biomedical and Life SciencesLancaster UniversityLancasterUK
  2. 2.NIHR Liverpool Pancreas Biomedical Research Unit, Department of Molecular and Clinical Cancer MedicineRoyal Liverpool University HospitalLiverpoolUK

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