Overexpression of MRP3 in HeLa-UGT1A9 Cells Enhances Glucuronidation Capability of the Cells
Copyright© Bentham Science Publishers; For any queries, please email at epubbenthamscience.net..
BACKGROUND: The interplay between phase II enzymes and efflux transporters leads to extensive metabolism and low systemic bioavailability of flavonoids.
OBJECTIVE: In this study, the dynamic interplay between multiple UGTs and multiple efflux transporters that occur inside the cells was fully investigated.
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 fraction of genistein metabolized (fmet) increased (14%, p<0.01) in the new cells. Small interfering (siRNA)-mediated MRP3 functional knockdown resulted in marked decreases in the excretion rates (26%-78%), intracellular amounts (56%-93%), and cellular clearance (54%-96%) in both cells, but the magnitude of the differences in HeLa- UGT1A9-Con cells was 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 as well as UGT mRNA expression suggest that HeLa-UGT1A9-MRP3 has increased expression of both MRP3 and UGT1A3.
CONCLUSION: The newly engineered HeLa-UGT1A9-MRP3 cells is an appropriate model to study the kinetic interplay between multiple UGTs and efflux transporters, and a promising biosynthetic tool to obtain flavonoid glucuronides of high purity.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:22 |
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| Enthalten in: |
Current drug metabolism - 22(2021), 10 vom: 09., Seite 772-783 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Zhou, Qiong [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 28.02.2022 Date Revised 28.02.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.2174/1389200222666210716151520 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM328245488 |
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| 245 | 1 | 0 | |a Overexpression of MRP3 in HeLa-UGT1A9 Cells Enhances Glucuronidation Capability of the Cells |
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| 500 | |a Date Revised 28.02.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright© Bentham Science Publishers; For any queries, please email at epubbenthamscience.net. | ||
| 520 | |a BACKGROUND: The interplay between phase II enzymes and efflux transporters leads to extensive metabolism and low systemic bioavailability of flavonoids | ||
| 520 | |a OBJECTIVE: In this study, the dynamic interplay between multiple UGTs and multiple efflux transporters that occur inside the cells was fully investigated | ||
| 520 | |a 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) | ||
| 520 | |a RESULTS: The excretion rate grew nearly 6-fold, cellular clearance of glucuronides increased about 3-fold, and fraction of genistein metabolized (fmet) increased (14%, p<0.01) in the new cells. Small interfering (siRNA)-mediated MRP3 functional knockdown resulted in marked decreases in the excretion rates (26%-78%), intracellular amounts (56%-93%), and cellular clearance (54%-96%) in both cells, but the magnitude of the differences in HeLa- UGT1A9-Con cells was 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 as well as UGT mRNA expression suggest that HeLa-UGT1A9-MRP3 has increased expression of both MRP3 and UGT1A3 | ||
| 520 | |a CONCLUSION: The newly engineered HeLa-UGT1A9-MRP3 cells is an appropriate model to study the kinetic interplay between multiple UGTs and efflux transporters, and a promising biosynthetic tool to obtain flavonoid glucuronides of high purity | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a HeLa cells. | |
| 650 | 4 | |a Multidrug resistance protein 3 (MRP3) | |
| 650 | 4 | |a UGT | |
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| 650 | 4 | |a genistein | |
| 650 | 4 | |a interplay | |
| 650 | 7 | |a ABCG2 protein, human |2 NLM | |
| 650 | 7 | |a ATP Binding Cassette Transporter, Subfamily G, Member 2 |2 NLM | |
| 650 | 7 | |a Flavonoids |2 NLM | |
| 650 | 7 | |a Multidrug Resistance-Associated Proteins |2 NLM | |
| 650 | 7 | |a Neoplasm Proteins |2 NLM | |
| 650 | 7 | |a multidrug resistance-associated protein 3 |2 NLM | |
| 650 | 7 | |a 1YV0492L5Z |2 NLM | |
| 650 | 7 | |a Genistein |2 NLM | |
| 650 | 7 | |a DH2M523P0H |2 NLM | |
| 650 | 7 | |a UDP-Glucuronosyltransferase 1A9 |2 NLM | |
| 650 | 7 | |a EC 2.4.1.17 |2 NLM | |
| 700 | 1 | |a Xia, Bijun |e verfasserin |4 aut | |
| 700 | 1 | |a Yin, Taijun |e verfasserin |4 aut | |
| 700 | 1 | |a He, Yu |e verfasserin |4 aut | |
| 700 | 1 | |a Ye, Ling |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Ming |e verfasserin |4 aut | |
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