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Overexpression of MRP3 in HeLa-UGT1A9 Cells Enhances Glucuronidation Capability of the Cells

Author(s):

Qiong Zhou, Bijun Xia, Taijun Yin, Yu He, Ling Ye and Ming Hu*   Pages 1 - 12 ( 12 )

Abstract:


Background. The interplay between phase II enzymes and efflux transporters leads to extensive metabolism and low systemic bioavailability of flavonoids.

Objective. The study aims to investigate the dynamic interplay between multiple UGTs and multiple efflux transporters inside the cells.

Methods. A new HeLa-UGT1A9-MRP3 cell was established to overexpress two dominant efflux transporters MRP3 and BCRP, and two UGT isoforms UGT1A9 and UGT1A3. The metabolism and glucuronides excretion for a model flavonoid genistein were determined in HeLa-UGT1A9-MRP3 cells and HeLa-UGT1A9-Con cells that overexpressed one UGT (1A9) and one efflux transporter (BCRP).

Results. The excretion rate grew nearly 6-fold, cellular clearance of glucuronides increased about 3-fold, and a fraction of genistein metabolized (fmet) increased (14%, p<0.01) in the new cells. Small interfering (siRNA)-mediated MRP3 functional knockdown resulted in markedly decreases in the excretion rates (26%-78%), intracellular amounts (56%-93%), cellular clearance (54%-96%) in both cells, but the magnitude of the differences in HeLa-UGT1A9-Con cells were relatively small. Reductions in fmet values were similarly moderate (11%-14%). In contrast, UGT1A9 knockdown with siRNA caused large decreases in the excretion rates (46%-88%), intracellular amounts (80%-97%), cellular clearance (80%-98%) as well as fmet value (33%-43%, p<0.01) in both UGT1A9 cells. Comparisons of the kinetic parameters and profiles of genistein glucuronidation and UGT mRNA expression suggest that HeLa-UGT1A9-MRP3 has increased expression of both MRP3 and UGT1A3.

Conclusion. The newly engineered HeLa-UGT1A9-MRP3 cells are an appropriate model to study the kinetic interplay between multiple UGTs and efflux transporters. It's a promising biosynthetic tool to obtain flavonoids glucuronides of high purity.

Keywords:

Multidrug resistance protein 3 (MRP3), UGT, Interplay, Flavonoid, Genistein, HeLa cells

Affiliation:

Department of Pharmacy, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, 4849 Calhoun Road, Houston, TX77204, Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, 4849 Calhoun Road, Houston, TX77204, Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, 4849 Calhoun Road, Houston, TX77204, Department of Pharmaceutics, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, 4849 Calhoun Road, Houston, TX77204



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