Characterization of P-glycoprotein and multidrug resistance proteins in rat kidney and intestinal cell lines

Water, Femke M. van de; Boleij, Annemarie; Peters, Janny G. P.; Rüssel, Frans G. M.; Masereeuw, Rosalinde

doi:10.1016/j.ejps.2006.09.008

Cited by 24 publications

(14 citation statements)

References 40 publications

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“…The selection of cell lines used in these experiments was based on the rate of cell growth, since cell confluency influences P-gp activity [8]. All cells were plated at similar densities at experimental start day -1, cultured overnight at 33 °C, and transferred to 37 °C at day 0, starting maturation.…”

Section: Resultsmentioning

confidence: 99%

Role of P-Glycoprotein Expression and Function in Cystinotic Renal Proximal Tubular Cells

et al. 2011

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P-glycoprotein (P-gp) is an ATP-dependent transporter localized at the apical membrane of the kidney proximal tubules, which plays a role in the efflux of cationic and amphipathic endogenous waste products and xenobiotics, such as drugs, into urine. Studies in mice deficient in P-gp showed generalized proximal tubular dysfunction similar to the phenotype of patients with cystinosis, an autosomal recessive disorder caused by mutations in the lysosomal cystine transporter cystinosin. Renal disease in cystinosis is characterized by generalized dysfunction of the apical proximal tubular influx transporters (so-called renal Fanconi syndrome) developing during infancy and gradually progressing towards end-stage renal disease before the 10th birthday in the majority of patients that are not treated with the cystine-depleting drug cysteamine. Here, we investigated whether the proximal tubular efflux transporter P-gp is affected in cystinosis and whether this might contribute to the development of renal Fanconi syndrome. We used conditionally immortalized (ci) proximal tubular epithelial cells (ciPTEC) derived from cystinotic patients and healthy volunteers. P-gp-mediated transport was measured by using the P-gp substrate calcein-AM in the presence and absence of the P-gp-inhibitor PSC833. P-gp activity was normal in cystinotic cells as compared to controls. Additionally, the effect of cysteamine on P-gp transport activity and phosphate uptake was determined; demonstrating increased P-gp activity in cystinotic cells, and further decrease of proximal tubular phosphate uptake. This observation is compatible with the persistence of renal Fanconi syndrome in vivo under cysteamine therapy. In summary, P-gp expression and activity are normal in cystinotic ciPTEC, indicating that P-gp dysfunction is not involved in the pathogenesis of cystinosis.

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Section: Resultsmentioning

confidence: 99%

Role of P-Glycoprotein Expression and Function in Cystinotic Renal Proximal Tubular Cells

et al. 2011

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“…So, we next decided to evaluate the effect of ABC transport inhibitors on the influx and efflux of 14 C-BT in a noncarcinogenic cell line, the IEC-6 cells. These cells express MDR1, MRP1, MRP3, MRP4, and MRP5, and show MDR1-and MRPs-mediated transport (40,64). Uptake of 14 C-BT by IEC-6 cells was not affected by verapamil or vinblastine.…”

Section: Effect Of Inhibitors Of Abc Transporters On Influx and Efflumentioning

confidence: 90%

“…The Caco-2 cell line was obtained from the Deutsche Sammlung von Mikroorganismen and Zellkulturen (Braunschweig, Germany) and was used between passage numbers [53][54][55][56][57][58][59][60][61][62][63][64][65][66][67][68][69][70][71]. The cells were maintained in a humidified atmosphere of 5% CO 2-95% air and were grown in Minimum Essential Medium (Sigma, St. Louis, MO) containing 5.55 mM glucose and supplemented with 15% fetal calf serum, 25 mM HEPES, 100 U/ml penicillin, 100 g/ml streptomycin, and 0.25 g/ml amphotericin B (all from Sigma).…”

Section: Caco-2 Cell Culturementioning

confidence: 99%

The short-chain fatty acid butyrate is a substrate of breast cancer resistance protein

Gonçalves

Gregório

Martel

2011

American Journal of Physiology-Cell Physiology

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Gonçalves P, Gregório I, Martel F. The short-chain fatty acid butyrate is a substrate of breast cancer resistance protein. Am J Physiol Cell Physiol 301: C984 -C994, 2011. First published July 20, 2011 doi:10.1152/ajpcell.00146.2011.-Colorectal cancer is one of the most common cancers worldwide. Butyrate (BT) plays a key role in colonic epithelium homeostasis. The aim of this work was to investigate the possibility of BT being transported by P-glycoprotein (MDR1), multidrug resistance proteins (MRPs), or breast cancer resistance protein (BCRP). Uptake and efflux of 14 C-BT and 3 H-folic acid were measured in Caco-2, IEC-6, and MDA-MB-231 cell lines. mRNA expression of BCRP was detected by RT-PCR. Cell viability, proliferation, and differentiation were quantified with the lactate dehydrogenase, sulforhodamine B, and alkaline phosphatase activity assays, respectively. In both IEC-6 cells and Caco-2 cells, no evidence was found for the involvement of either MDR1 or MRPs in 14 C-BT efflux from the cells. In contrast, several lines of evidence support the conclusion that BT is a substrate of both rat and human BCRP. Indeed, BCRP inhibitors reduced 14 C-BT efflux in IEC-6 cells, both BT and BCRP inhibitors significantly decreased the efflux of the known BCRP substrate 3 H-folic acid in IEC-6 cells, and BCRP inhibitors reduced 14 C-BT efflux in the BCRP-expressing MDA-MB-231 cell line. In IEC-6 cells, combination of BT with a BCRP inhibitor significantly potentiated the effect of BT on cell proliferation. The results of this study, showing for the first time that BT is a BCRP substrate, are very important in the context of the high levels of BCRP expression in the human colon and the anticarcinogenic and anti-inflammatory role of BT at that level. So, interaction of BT with BCRP and with other BCRP substrates/inhibitors is clearly of major importance. butyrate efflux; colorectal cancer; IEC-6 cells; anticarcinogenic effect COLORECTAL CANCER (CRC) is a leading cause of cancer death in occidental countries (36). Butyrate (BT), a product of intestinal flora fermentation of dietary fiber, has a protective role in the prevention and progression of colorectal carcinogenesis (57). Indeed, this short-chain fatty acid plays a key role in colonic epithelium homeostasis, by having multiple important roles at that level: 1) it is the main energy source for colonocytes, 2) it promotes growth and proliferation of normal colonic epithelial cells, 3) it inhibits colon carcinogenesis (by suppressing growth of cancer cells, inducing differentiation and apoptosis and inhibiting cell proliferation), 4) it inhibits colon inflammation and oxidative stress, 5) it improves the colonic defence barrier function, and 6) it stimulates fluid and electrolyte absorption (reviews in Refs. 31, 68). 1The mechanism by which BT inhibits colon carcinogenesis seems to involve various effects on gene expression, which are mainly attributed to its capacity to act as a histone deacetylase inhibitor (HDAC), leading to hyperacetylation of histones and to incr...

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“…The activity of the ABC efflux transporter Pgp was assessed by measuring the accumulation of calcein as described before (van de Water et al 2007). Briefly, matured cells were incubated in two independent experiments for 1 h at 37°C with the lipophylic non-fluorescent Pgp substrate calcein-AM (Invitrogen) in the presence or absence of inhibitor PSC-833, which was a kind gift from Novartis Pharma (Basel, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

Novel conditionally immortalized human proximal tubule cell line expressing functional influx and efflux transporters

et al. 2009

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Reabsorption of filtered solutes from the glomerular filtrate and excretion of waste products and xenobiotics are the main functions of the renal proximal tubular (PT) epithelium. A human PT cell line expressing a range of functional transporters would help to augment current knowledge in renal physiology and pharmacology. We have established and characterized a conditionally immortalized PT epithelial cell line (ciPTEC) obtained by transfecting and subcloning cells exfoliated in the urine of a healthy volunteer. The PT origin of this line has been confirmed morphologically and by the expression of aminopeptidase N, zona occludens 1, aquaporin 1, dipeptidyl peptidase IV and multidrug resistance protein 4 together with alkaline phosphatase activity. ciPTEC assembles in a tight monolayer with limited diffusion of inulin-fluorescein-isothiocyanate. Concentration and time-dependent reabsorption of albumin via endocytosis has been demonstrated, together with sodium-dependent phosphate uptake. The expression and activity of apical efflux transporter p-glycoprotein and of baso-lateral influx transporter organic cation transporter 2 have been shown in ciPTEC. This established human ciPTEC expressing multiple endogenous organic ion transporters mimicking renal reabsorption and excretion represents a powerful tool for future in vitro transport studies in pharmacology and physiology.

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Characterization of P-glycoprotein and multidrug resistance proteins in rat kidney and intestinal cell lines

Cited by 24 publications

References 40 publications

Role of P-Glycoprotein Expression and Function in Cystinotic Renal Proximal Tubular Cells

Role of P-Glycoprotein Expression and Function in Cystinotic Renal Proximal Tubular Cells

The short-chain fatty acid butyrate is a substrate of breast cancer resistance protein

Novel conditionally immortalized human proximal tubule cell line expressing functional influx and efflux transporters

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