P-Glycoprotein expression in human retinal pigment epithelium cell lines

Experimental Eye Research
Paul A ConstableN Joan Abbott

Abstract

P-Glycoprotein (P-gp), an active efflux transporter encoded by the MDR1 gene, has recently been identified in the human and pig retinal pigment epithelium (RPE) in situ. Efflux pumps such as P-gp are major barriers to drug delivery in several tissues. We wished to establish whether human RPE cell lines express P-gp under the culture conditions recommended for each cell line so as to determine their suitability as in vitro models for predicting drug transport across the outer blood-retinal barrier. Three human RPE cell lines, ARPE19, D407 and h1RPE were investigated. Reverse transcriptase-polymerase chain reaction (RT-PCR) was carried out to determine the expression of MDR1 mRNA. Immunocytochemistry using the P-gp-specific antibody C219 was undertaken to investigate the presence of P-gp protein in each cell type. Uptake of rhodamine 123, a P-gp substrate, in the presence or absence of pre-treatment with a P-gp inhibitor, verapamil, was measured in each cell line to determine functional expression of P-gp. For all experiments, MDCK cells stably transfected with the human MDR1 gene (MDCK-MDR1) were used as a positive control. ARPE19 cells were consistently negative for P-gp as assessed by RT-PCR and immunocytochemistry. By contras...Continue Reading

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Citations

Sep 2, 2008·Pharmaceutical Research·Ripal GaudanaAshim K Mitra
Apr 23, 2009·Pharmaceutical Research·Eliisa MannermaaArto Urtti
May 15, 2008·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Srinivasan SenthilkumariSupriyo Ghose
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Sep 2, 2014·Journal of Ophthalmology·Alla V KuznetsovaMaria A Aleksandrova
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Feb 9, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Eun-Jeong ChoiHea-Young Cho
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Jul 2, 2021·Progress in Biophysics and Molecular Biology·Ivana Barravecchia, Gian Carlo Demontis

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