UDP-Glucuronosyltransferase (UGT) 1A9-Overexpressing HeLa Cells Is an Appropriate Tool to Delineate the Kinetic Interplay between Breast Cancer Resistance Protein (BRCP) and UGT and to Rapidly Identify the Glucuronide Substrates of BCRP

Jiang, Wen Qi; Xu, Beibei; Wu, Baojian; Yu, Rong; Hu, Ming

doi:10.1124/dmd.111.041467

Cited by 37 publications

(52 citation statements)

References 34 publications

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“…We also demonstrated that cellular excretion of CG was contributed by multiple efflux transporters (including BCRP, MRP1, MRP3, and MRP4) using a combined approach of chemical inhibition and shRNA-mediated silencing. Our findings were consistent with the study of Jiang et al (2012) in which BCRP was involved in excretion of flavonoid glucuronides in HeLa cells. However, Jiang et al (2012) showed that contributions of MRP family proteins (e.g., MRP2/ MRP3) to glucuronide excretion were negligible.…”

Section: Discussionsupporting

confidence: 82%

“…Our findings were consistent with the study of Jiang et al (2012) in which BCRP was involved in excretion of flavonoid glucuronides in HeLa cells. However, Jiang et al (2012) showed that contributions of MRP family proteins (e.g., MRP2/ MRP3) to glucuronide excretion were negligible. In our study, we provided strong evidence that MRP family proteins contributed significantly to excretion of CG.…”

Section: Discussionsupporting

confidence: 82%

“…LTC4 is a high-affinity substrate of MRP1/MRP2 (K m = 0.1-1 mM) that has been used to inhibit the transport of drugs/metabolites by MRPs (Loe et al, 1996;Cui et al, 1999;Miller et al, 2000;Hu et al, 2003;Jiang et al, 2012). However, the inhibition selectivity of LTC4 toward MRP family members was unknown.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide

et al. 2015

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Active transport of glucuronide out of cells is a critical process in elimination of drugs via the glucuronidation pathway. Here, HeLa cells were stably transfected with UGT1A1 and the contributions of BCRP and MRP family transporters to the cellular efflux of chrysin glucuronide (CG) were determined. The cDNA of UGT1A1 was introduced into HeLa cells using the lentiviral transfection method. The modified cells were functional in generation of the glucuronide from chrysin. Ko143 at 10-20 mM (a dual inhibitor of BCRP and UGT1A1) caused a marked decrease (51.3%-59.7%, P < 0.01) in the excretion rate and efflux clearance of CG. Likewise, MK-571 at 5-20 mM (an inhibitor of MRPs but an activator of UGT1A1) resulted in a significant reduction in the excretion rate (18.2%-64.0%, P < 0.01) and efflux clearance (37.0%-90.2%, P < 0.001). By contrast, dipyridamole and leukotriene C4 showed no inhibitory effects on CG excretion. The chemical inhibition indicated that excretion of CG was contributed by the MRP family transporters, whereas the role of BCRP was unclear. Furthermore, short hairpin RNA-mediated silencing of a target transporter led to a marked reduction in the excretion rate of CG (38.6% for BCRP, 39.3% for MRP1, 36.4% for MRP3, and 28.7% for MRP4; P < 0.01). Transporter silencing also led to substantial decreases in the efflux clearance (44.7% for BCRP, 60.4% for MRP1, 36.7% for MRP3, and 28.7% for MRP4; P < 0.01). The gene silencing results suggested that BCRP, MRP1, MRP3, and MRP4 were significant contributors to excretion of CG.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionsupporting

confidence: 82%

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Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide

et al. 2015

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“…The data points in figures are observed glucuronidation rates, and the curves are plotted using the fitted kinetic parameters (Table 2). MRP2 and BCRP are important transporters for the efflux of glucuronides (Jiang et al, 2012). We found that the amount of calycosin-39-glucuronide in the perfusate samples significantly decreased in the presence of MK571 and ko143 (Fig.…”

Section: Discussionmentioning

confidence: 67%

SGLT-1 Transport and Deglycosylation inside Intestinal Cells Are Key Steps in the Absorption and Disposition of Calycosin-7-O- -D-Glucoside in Rats

Shi

Zheng

et al. 2015

Drug Metabolism and Disposition

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Hydrolysis by lactase-phloridzin hydrolase (LPH) is the first and critical step in the absorption of isoflavonoid glucosides. However, the absorption characteristics of calycosin-7-O-b-D-glucoside (CG) slightly differ from other isoflavonoid glucosides. In this study, we used the rat intestinal perfusion model and performed pharmacokinetic studies and in vitro experiments to determine the factors influencing CG absorption and disposition. After oral administration of isoflavonoid glucosides, LPH was found to play minimal or no role on the hydrolysis of CG, in contrast to that of daidzin. CG was mainly transported into the small intestinal cells by sodium-dependent glucose transporter 1 (SGLT-1) as intact. This pathway could be the main mechanism underlying the high permeability of CG in the small intestine. CG was likely to be hydrolyzed in enterocytes to its aglycone calycosin by broad-specific b-glucuronides (BSbG) and glucocerebrosidase or rapidly metabolized. Calycosin was also rapidly and extensively metabolized to 39-glucuronide in the enterocytes and liver, and the glucuronidation rates of calycosin and CG were much higher in the former. The metabolites were also transported into lumen by breast cancer resistance protein and multidrug resistance-associated protein 2. In conclusion, the enterocytes could be an important site for CG absorption, deglycosylation, and metabolism in rats. This study could contribute to the theoretical foundation and mechanism of absorption and disposition of flavonoid compounds.

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“…A better understanding of transporter-enzyme interplay contributes to improved predictions of in vivo drug disposition and drug-drug interactions (Benet et al, 2003;Lam et al, 2006). In addition to P-glycoprotein/cytochrome p450 3A, the interplay is also applicable to the efflux transporters (i.e., breast cancer resistance protein and MRPs) and phase II enzymes (Jiang et al, 2012;Zhang et al, 2015). The dependence of phase II metabolism on efflux transporters is underpinned by the fact that the hydrophilic phase II metabolites (lacking in passive transport ability) require the efflux transporters for cellular excretion (Wu, 2012).…”

Section: Introductionmentioning

confidence: 99%

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

Zhao¹,

Wang²,

Li³

et al. 2016

Drug Metabolism and Disposition

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Elucidating the intricate relationships between metabolic and transport pathways contributes to improved predictions of in vivo drug disposition and drug-drug interactions. Here we reported that inhibited excretion of conjugative metabolites [i.e., hesperetin 39-O-sulfate (H39S) and hesperetin 7-O-sulfate (H7S)] by MK-571 led to reduced metabolism of hesperetin (a maximal 78% reduction) in human embryonic kidney 293 cells overexpressing sulfotransferase 1A3 (named SULT293 cells). The strong dependence of cellular sulfonation on the efflux transport of generated sulfated metabolites revealed an interplay of sulfonation metabolism with efflux transport (or sulfonation-transport interplay). Polymerase chain reaction (PCR) and Western blot analyses demonstrated that SULT293 cells expressed multiple sulfatases such as arylsulfatase A (ARSA), ARSB, and ARSC. Of these three desulfonation enzymes, only ARSB showed significant activities toward hesperetin sulfates. The intrinsic clearance values for the hydrolysis of H39S and H7S were estimated at 0.6 and 0.5 ml/h/mg, respectively. Furthermore, knockdown of ARSB attenuated the regulatory effect of efflux transporter on cellular sulfonation, whereas overexpression of ABSB enhanced the transporter effect. Taken together, the results indicated that ARSB mediated the sulfonation-transport interplay in SULT293 cells.

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UDP-Glucuronosyltransferase (UGT) 1A9-Overexpressing HeLa Cells Is an Appropriate Tool to Delineate the Kinetic Interplay between Breast Cancer Resistance Protein (BRCP) and UGT and to Rapidly Identify the Glucuronide Substrates of BCRP

Cited by 37 publications

References 34 publications

Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide

Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide

SGLT-1 Transport and Deglycosylation inside Intestinal Cells Are Key Steps in the Absorption and Disposition of Calycosin-7-O- -D-Glucoside in Rats

Arylsulfatase B Mediates the Sulfonation-Transport Interplay in Human Embryonic Kidney 293 Cells Overexpressing Sulfotransferase 1A3

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