Regulation of the expression of renal drug transporters in KEAP1-knockdown human tubular cells
•The KEAP1 knockdown elevated several ABC transporters expression in human tubular epithelial HK-2.•The expression of MRP2-3, MDR1, and BCRP were enhanced by genetic activation of NRF2.•The KEAP1 knockdown HK-2 was resistant to drug accumulation and anticancer drug cytotoxicity.•The pharmacological...
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| Veröffentlicht in: | Toxicology in vitro 2015-08, Vol.29 (5), p.884-892 |
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| creator | Jeong, Hyuk-Sang Ryoo, In-geun Kwak, Mi-Kyoung |
| description | •The KEAP1 knockdown elevated several ABC transporters expression in human tubular epithelial HK-2.•The expression of MRP2-3, MDR1, and BCRP were enhanced by genetic activation of NRF2.•The KEAP1 knockdown HK-2 was resistant to drug accumulation and anticancer drug cytotoxicity.•The pharmacological NRF2 activation did not show noticeable changes in ABC transporters expression.
The kidney secretes various xenobiotics through a well-established transport system. The transcription factor NF-E2-related factor 2 (NRF2) up-regulates a subset of genes encoding antioxidant and detoxification proteins. Kelch-like ECH-associated protein 1 (KEAP1) down-regulates NRF2 by facilitating continuous degradation of NRF2 protein. Here, we investigated the role of NRF2 in the expression of renal drug transporters by using a stable KEAP1 knockdown renal tubular HK-2 cell line (shKEAP1). KEAP1 knockdown resulted in a significant increase in the expression of four renal transporters, namely, multidrug resistance protein 1 (MDR1; ABCB1), breast cancer resistance protein (BCRP; ABCG2), multidrug resistance-associated protein 2 (MRP2; ABCC2), and MRP3 (ABCC3). In western blot and immunocytochemical analyses, protein levels of these transporters were also significantly higher in the knockdown group. Consequently, shKEAP1 cells released more Hoechst 33342 fluorescent dye and doxorubicin, and they were more resistant to doxorubicin than the control cells. In addition, cisplatin resistance of shKEAP1 decreased upon co-incubation with a transporter inhibitor. Whereas, a short term incubation (24h) with sulforaphane did not show noticeable changes in the expression of transporter. Collectively, these results indicate that NRF2 regulates the expression of MDR1, BCRP, MRP2, and MRP3 in human tubular epithelial cells. Altered expression of these transporters affects drug secretion in these cells, which may result in the renal cellular damage upon exposure to nephrotoxic xenobiotics. |
| doi_str_mv | 10.1016/j.tiv.2015.03.013 |
| format | Article |
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The kidney secretes various xenobiotics through a well-established transport system. The transcription factor NF-E2-related factor 2 (NRF2) up-regulates a subset of genes encoding antioxidant and detoxification proteins. Kelch-like ECH-associated protein 1 (KEAP1) down-regulates NRF2 by facilitating continuous degradation of NRF2 protein. Here, we investigated the role of NRF2 in the expression of renal drug transporters by using a stable KEAP1 knockdown renal tubular HK-2 cell line (shKEAP1). KEAP1 knockdown resulted in a significant increase in the expression of four renal transporters, namely, multidrug resistance protein 1 (MDR1; ABCB1), breast cancer resistance protein (BCRP; ABCG2), multidrug resistance-associated protein 2 (MRP2; ABCC2), and MRP3 (ABCC3). In western blot and immunocytochemical analyses, protein levels of these transporters were also significantly higher in the knockdown group. Consequently, shKEAP1 cells released more Hoechst 33342 fluorescent dye and doxorubicin, and they were more resistant to doxorubicin than the control cells. In addition, cisplatin resistance of shKEAP1 decreased upon co-incubation with a transporter inhibitor. Whereas, a short term incubation (24h) with sulforaphane did not show noticeable changes in the expression of transporter. Collectively, these results indicate that NRF2 regulates the expression of MDR1, BCRP, MRP2, and MRP3 in human tubular epithelial cells. Altered expression of these transporters affects drug secretion in these cells, which may result in the renal cellular damage upon exposure to nephrotoxic xenobiotics.</description><identifier>ISSN: 0887-2333</identifier><identifier>EISSN: 1879-3177</identifier><identifier>DOI: 10.1016/j.tiv.2015.03.013</identifier><identifier>PMID: 25841332</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>ABC transporters ; ATP-Binding Cassette Transporters - metabolism ; Benzimidazoles - pharmacology ; Cell Line ; Cisplatin - pharmacology ; Doxorubicin - pharmacology ; Gene Knockdown Techniques ; Humans ; Intracellular Signaling Peptides and Proteins - genetics ; Isothiocyanates - pharmacology ; KEAP1 ; Kelch-Like ECH-Associated Protein 1 ; NF-E2-Related Factor 2 - metabolism ; NRF2 ; Propionates - pharmacology ; Proximal tubule epithelial cells ; Quinolines - pharmacology ; SLC transporters ; Sulforaphane</subject><ispartof>Toxicology in vitro, 2015-08, Vol.29 (5), p.884-892</ispartof><rights>2015 Elsevier Ltd</rights><rights>Copyright © 2015 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c452t-daa02ccac456c06e9b3c7f186fc1a5c4cc6df30e0d66917417321d8bd325cc713</citedby><cites>FETCH-LOGICAL-c452t-daa02ccac456c06e9b3c7f186fc1a5c4cc6df30e0d66917417321d8bd325cc713</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.tiv.2015.03.013$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25841332$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jeong, Hyuk-Sang</creatorcontrib><creatorcontrib>Ryoo, In-geun</creatorcontrib><creatorcontrib>Kwak, Mi-Kyoung</creatorcontrib><title>Regulation of the expression of renal drug transporters in KEAP1-knockdown human tubular cells</title><title>Toxicology in vitro</title><addtitle>Toxicol In Vitro</addtitle><description>•The KEAP1 knockdown elevated several ABC transporters expression in human tubular epithelial HK-2.•The expression of MRP2-3, MDR1, and BCRP were enhanced by genetic activation of NRF2.•The KEAP1 knockdown HK-2 was resistant to drug accumulation and anticancer drug cytotoxicity.•The pharmacological NRF2 activation did not show noticeable changes in ABC transporters expression.
The kidney secretes various xenobiotics through a well-established transport system. The transcription factor NF-E2-related factor 2 (NRF2) up-regulates a subset of genes encoding antioxidant and detoxification proteins. Kelch-like ECH-associated protein 1 (KEAP1) down-regulates NRF2 by facilitating continuous degradation of NRF2 protein. Here, we investigated the role of NRF2 in the expression of renal drug transporters by using a stable KEAP1 knockdown renal tubular HK-2 cell line (shKEAP1). KEAP1 knockdown resulted in a significant increase in the expression of four renal transporters, namely, multidrug resistance protein 1 (MDR1; ABCB1), breast cancer resistance protein (BCRP; ABCG2), multidrug resistance-associated protein 2 (MRP2; ABCC2), and MRP3 (ABCC3). In western blot and immunocytochemical analyses, protein levels of these transporters were also significantly higher in the knockdown group. Consequently, shKEAP1 cells released more Hoechst 33342 fluorescent dye and doxorubicin, and they were more resistant to doxorubicin than the control cells. In addition, cisplatin resistance of shKEAP1 decreased upon co-incubation with a transporter inhibitor. Whereas, a short term incubation (24h) with sulforaphane did not show noticeable changes in the expression of transporter. Collectively, these results indicate that NRF2 regulates the expression of MDR1, BCRP, MRP2, and MRP3 in human tubular epithelial cells. Altered expression of these transporters affects drug secretion in these cells, which may result in the renal cellular damage upon exposure to nephrotoxic xenobiotics.</description><subject>ABC transporters</subject><subject>ATP-Binding Cassette Transporters - metabolism</subject><subject>Benzimidazoles - pharmacology</subject><subject>Cell Line</subject><subject>Cisplatin - pharmacology</subject><subject>Doxorubicin - pharmacology</subject><subject>Gene Knockdown Techniques</subject><subject>Humans</subject><subject>Intracellular Signaling Peptides and Proteins - genetics</subject><subject>Isothiocyanates - pharmacology</subject><subject>KEAP1</subject><subject>Kelch-Like ECH-Associated Protein 1</subject><subject>NF-E2-Related Factor 2 - metabolism</subject><subject>NRF2</subject><subject>Propionates - pharmacology</subject><subject>Proximal tubule epithelial cells</subject><subject>Quinolines - pharmacology</subject><subject>SLC transporters</subject><subject>Sulforaphane</subject><issn>0887-2333</issn><issn>1879-3177</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1PGzEQhi1UVALtD-BS-djLbj129iPqCSE-qiJRVXCt5YxnwWFjp7aXwr_HUdIee7LGet5XMw9jpyBqENB-WdXZPddSQFMLVQtQB2wGfbeoFHTdOzYTfd9VUil1xI5TWgkhml6K9-xINv0clJIz9usnPUyjyS54HgaeH4nTyyZSSvufSN6M3MbpgedofNqEmCkm7jz_fnH2A6onH_DJhj-eP05r43melqUwcqRxTB_Y4WDGRB_37wm7v7y4O7-ubm6vvp2f3VQ4b2SurDFCIpoytShaWiwVdgP07YBgGpwjtnZQgoRt2wV0c-iUBNsvrZINYgfqhH3e9W5i-D1Rynrt0nYD4ylMSZdAswApZVNQ2KEYQ0qRBr2Jbm3iqwaht1r1SheteqtVC6WL1pL5tK-flmuy_xJ_PRbg6w6gcuSzo6gTOvJI1kXCrG1w_6l_A-oLiTw</recordid><startdate>201508</startdate><enddate>201508</enddate><creator>Jeong, Hyuk-Sang</creator><creator>Ryoo, In-geun</creator><creator>Kwak, Mi-Kyoung</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U7</scope><scope>C1K</scope></search><sort><creationdate>201508</creationdate><title>Regulation of the expression of renal drug transporters in KEAP1-knockdown human tubular cells</title><author>Jeong, Hyuk-Sang ; Ryoo, In-geun ; Kwak, Mi-Kyoung</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c452t-daa02ccac456c06e9b3c7f186fc1a5c4cc6df30e0d66917417321d8bd325cc713</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>ABC transporters</topic><topic>ATP-Binding Cassette Transporters - metabolism</topic><topic>Benzimidazoles - pharmacology</topic><topic>Cell Line</topic><topic>Cisplatin - pharmacology</topic><topic>Doxorubicin - pharmacology</topic><topic>Gene Knockdown Techniques</topic><topic>Humans</topic><topic>Intracellular Signaling Peptides and Proteins - genetics</topic><topic>Isothiocyanates - pharmacology</topic><topic>KEAP1</topic><topic>Kelch-Like ECH-Associated Protein 1</topic><topic>NF-E2-Related Factor 2 - metabolism</topic><topic>NRF2</topic><topic>Propionates - pharmacology</topic><topic>Proximal tubule epithelial cells</topic><topic>Quinolines - pharmacology</topic><topic>SLC transporters</topic><topic>Sulforaphane</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jeong, Hyuk-Sang</creatorcontrib><creatorcontrib>Ryoo, In-geun</creatorcontrib><creatorcontrib>Kwak, Mi-Kyoung</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Toxicology Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Toxicology in vitro</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jeong, Hyuk-Sang</au><au>Ryoo, In-geun</au><au>Kwak, Mi-Kyoung</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Regulation of the expression of renal drug transporters in KEAP1-knockdown human tubular cells</atitle><jtitle>Toxicology in vitro</jtitle><addtitle>Toxicol In Vitro</addtitle><date>2015-08</date><risdate>2015</risdate><volume>29</volume><issue>5</issue><spage>884</spage><epage>892</epage><pages>884-892</pages><issn>0887-2333</issn><eissn>1879-3177</eissn><abstract>•The KEAP1 knockdown elevated several ABC transporters expression in human tubular epithelial HK-2.•The expression of MRP2-3, MDR1, and BCRP were enhanced by genetic activation of NRF2.•The KEAP1 knockdown HK-2 was resistant to drug accumulation and anticancer drug cytotoxicity.•The pharmacological NRF2 activation did not show noticeable changes in ABC transporters expression.
The kidney secretes various xenobiotics through a well-established transport system. The transcription factor NF-E2-related factor 2 (NRF2) up-regulates a subset of genes encoding antioxidant and detoxification proteins. Kelch-like ECH-associated protein 1 (KEAP1) down-regulates NRF2 by facilitating continuous degradation of NRF2 protein. Here, we investigated the role of NRF2 in the expression of renal drug transporters by using a stable KEAP1 knockdown renal tubular HK-2 cell line (shKEAP1). KEAP1 knockdown resulted in a significant increase in the expression of four renal transporters, namely, multidrug resistance protein 1 (MDR1; ABCB1), breast cancer resistance protein (BCRP; ABCG2), multidrug resistance-associated protein 2 (MRP2; ABCC2), and MRP3 (ABCC3). In western blot and immunocytochemical analyses, protein levels of these transporters were also significantly higher in the knockdown group. Consequently, shKEAP1 cells released more Hoechst 33342 fluorescent dye and doxorubicin, and they were more resistant to doxorubicin than the control cells. In addition, cisplatin resistance of shKEAP1 decreased upon co-incubation with a transporter inhibitor. Whereas, a short term incubation (24h) with sulforaphane did not show noticeable changes in the expression of transporter. Collectively, these results indicate that NRF2 regulates the expression of MDR1, BCRP, MRP2, and MRP3 in human tubular epithelial cells. Altered expression of these transporters affects drug secretion in these cells, which may result in the renal cellular damage upon exposure to nephrotoxic xenobiotics.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>25841332</pmid><doi>10.1016/j.tiv.2015.03.013</doi><tpages>9</tpages></addata></record> |
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| subjects | ABC transporters ATP-Binding Cassette Transporters - metabolism Benzimidazoles - pharmacology Cell Line Cisplatin - pharmacology Doxorubicin - pharmacology Gene Knockdown Techniques Humans Intracellular Signaling Peptides and Proteins - genetics Isothiocyanates - pharmacology KEAP1 Kelch-Like ECH-Associated Protein 1 NF-E2-Related Factor 2 - metabolism NRF2 Propionates - pharmacology Proximal tubule epithelial cells Quinolines - pharmacology SLC transporters Sulforaphane |
| title | Regulation of the expression of renal drug transporters in KEAP1-knockdown human tubular cells |
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