A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis
Background Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mi...
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| Veröffentlicht in: | Cytometry. Part B, Clinical cytometry Clinical cytometry, 2004-05, Vol.59B (1), p.46-53 |
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| container_title | Cytometry. Part B, Clinical cytometry |
| container_volume | 59B |
| creator | Barbier, M. Gray, B. D. Muirhead, K. A. Ronot, X. Boutonnat, J. |
| description | Background
Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mitochondrial potential, in chemosensitive and chemoresistant tumor cell populations.
Methods
A three‐color labeling was performed using Hoechst 33342 (DNA), JC1 (mitochondrial potential), and a far red fluorescent membrane intercalating dye: PTIR271 (proliferation).
Results
Combined staining of DNA and mitochondrial potential allows identification of subpopulations expressing and MDR phenotype mediated by P‐glycoprotein (Pgp), and, in Pgp negative subpopulations, identification of apoptotic cells and evaluation of cell cycle status in viable cells. Addition of a far red fluorescent membrane intercalating dye, PTIR271, allows simultaneous monitoring of cell division status by dye dilution in both drug sensitive and drug resistant populations.
Conclusion
This triple labeling is an interesting method to study the proliferation status of drug sensitivity and drug resistance in viable tumor cells. © 2004 Wiley‐Liss, Inc. |
| doi_str_mv | 10.1002/cyto.b.20004 |
| format | Article |
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Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mitochondrial potential, in chemosensitive and chemoresistant tumor cell populations.
Methods
A three‐color labeling was performed using Hoechst 33342 (DNA), JC1 (mitochondrial potential), and a far red fluorescent membrane intercalating dye: PTIR271 (proliferation).
Results
Combined staining of DNA and mitochondrial potential allows identification of subpopulations expressing and MDR phenotype mediated by P‐glycoprotein (Pgp), and, in Pgp negative subpopulations, identification of apoptotic cells and evaluation of cell cycle status in viable cells. Addition of a far red fluorescent membrane intercalating dye, PTIR271, allows simultaneous monitoring of cell division status by dye dilution in both drug sensitive and drug resistant populations.
Conclusion
This triple labeling is an interesting method to study the proliferation status of drug sensitivity and drug resistance in viable tumor cells. © 2004 Wiley‐Liss, Inc.</description><identifier>ISSN: 1552-4949</identifier><identifier>EISSN: 1552-4957</identifier><identifier>DOI: 10.1002/cyto.b.20004</identifier><identifier>PMID: 15108170</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Antineoplastic Agents, Phytogenic - pharmacokinetics ; Antineoplastic Agents, Phytogenic - pharmacology ; apoptosis ; Apoptosis - drug effects ; Camptothecin - analogs & derivatives ; Camptothecin - pharmacokinetics ; Camptothecin - pharmacology ; Cell Division - drug effects ; Drug Resistance, Neoplasm ; Flow Cytometry - methods ; Fluorescent Dyes ; Humans ; K562 Cells ; MDR ; PKH dyes, PTIR271 ; Predictive Value of Tests ; proliferation monitoring ; Staining and Labeling - methods</subject><ispartof>Cytometry. Part B, Clinical cytometry, 2004-05, Vol.59B (1), p.46-53</ispartof><rights>Copyright © 2004 Wiley‐Liss, Inc.</rights><rights>Copyright 2004 Wiley-Liss, Inc.</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4004-f265a264c390387716cc82afe55abbd34ff7aa8440e36240e01148dda1c1d3343</citedby><cites>FETCH-LOGICAL-c4004-f265a264c390387716cc82afe55abbd34ff7aa8440e36240e01148dda1c1d3343</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fcyto.b.20004$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fcyto.b.20004$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1416,1432,27922,27923,45572,45573,46407,46831</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15108170$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Barbier, M.</creatorcontrib><creatorcontrib>Gray, B. D.</creatorcontrib><creatorcontrib>Muirhead, K. A.</creatorcontrib><creatorcontrib>Ronot, X.</creatorcontrib><creatorcontrib>Boutonnat, J.</creatorcontrib><title>A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis</title><title>Cytometry. Part B, Clinical cytometry</title><addtitle>Cytometry B Clin Cytom</addtitle><description>Background
Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mitochondrial potential, in chemosensitive and chemoresistant tumor cell populations.
Methods
A three‐color labeling was performed using Hoechst 33342 (DNA), JC1 (mitochondrial potential), and a far red fluorescent membrane intercalating dye: PTIR271 (proliferation).
Results
Combined staining of DNA and mitochondrial potential allows identification of subpopulations expressing and MDR phenotype mediated by P‐glycoprotein (Pgp), and, in Pgp negative subpopulations, identification of apoptotic cells and evaluation of cell cycle status in viable cells. Addition of a far red fluorescent membrane intercalating dye, PTIR271, allows simultaneous monitoring of cell division status by dye dilution in both drug sensitive and drug resistant populations.
Conclusion
This triple labeling is an interesting method to study the proliferation status of drug sensitivity and drug resistance in viable tumor cells. © 2004 Wiley‐Liss, Inc.</description><subject>Antineoplastic Agents, Phytogenic - pharmacokinetics</subject><subject>Antineoplastic Agents, Phytogenic - pharmacology</subject><subject>apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>Camptothecin - analogs & derivatives</subject><subject>Camptothecin - pharmacokinetics</subject><subject>Camptothecin - pharmacology</subject><subject>Cell Division - drug effects</subject><subject>Drug Resistance, Neoplasm</subject><subject>Flow Cytometry - methods</subject><subject>Fluorescent Dyes</subject><subject>Humans</subject><subject>K562 Cells</subject><subject>MDR</subject><subject>PKH dyes, PTIR271</subject><subject>Predictive Value of Tests</subject><subject>proliferation monitoring</subject><subject>Staining and Labeling - methods</subject><issn>1552-4949</issn><issn>1552-4957</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkDtPwzAURi0EoqWwMSNPTE3xK3EyVhUvCalLGZiM49goyImDnajk35OQCjZYfK2ro3M_fQBcYrTCCJEb1bdula8IQogdgTmOYxKxLObHP3-WzcBZCO8I0Zgl_BTMcIxRijmag9c1NNbt4WipdOtLBWUIsofGeRjKqrOtrLXrwrjWIVS6bqEzsPDdG9TG2O5zCRvvbGm0l23p6iWUdQFl45rWhTKcgxMjbdAXh7kAz3e3u81D9LS9f9ysnyLFhuCRIUksScIUzRBNOceJUimRRsexzPOCMmO4lCljSNOEDC_CmKVFIbHCBaWMLsD15B3CfHQ6tKIqg9LWTvEFx2nCM5T8C5LhPmYYD-ByApV3IXhtROPLSvpeYCTG6sVYmsjFd_UDfnXwdnmli1_40PUA0AnYl1b3f8rE5mW3nbRfXIaQ2g</recordid><startdate>200405</startdate><enddate>200405</enddate><creator>Barbier, M.</creator><creator>Gray, B. D.</creator><creator>Muirhead, K. A.</creator><creator>Ronot, X.</creator><creator>Boutonnat, J.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>200405</creationdate><title>A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis</title><author>Barbier, M. ; Gray, B. D. ; Muirhead, K. A. ; Ronot, X. ; Boutonnat, J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4004-f265a264c390387716cc82afe55abbd34ff7aa8440e36240e01148dda1c1d3343</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Antineoplastic Agents, Phytogenic - pharmacokinetics</topic><topic>Antineoplastic Agents, Phytogenic - pharmacology</topic><topic>apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>Camptothecin - analogs & derivatives</topic><topic>Camptothecin - pharmacokinetics</topic><topic>Camptothecin - pharmacology</topic><topic>Cell Division - drug effects</topic><topic>Drug Resistance, Neoplasm</topic><topic>Flow Cytometry - methods</topic><topic>Fluorescent Dyes</topic><topic>Humans</topic><topic>K562 Cells</topic><topic>MDR</topic><topic>PKH dyes, PTIR271</topic><topic>Predictive Value of Tests</topic><topic>proliferation monitoring</topic><topic>Staining and Labeling - methods</topic><toplevel>online_resources</toplevel><creatorcontrib>Barbier, M.</creatorcontrib><creatorcontrib>Gray, B. D.</creatorcontrib><creatorcontrib>Muirhead, K. A.</creatorcontrib><creatorcontrib>Ronot, X.</creatorcontrib><creatorcontrib>Boutonnat, J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Cytometry. Part B, Clinical cytometry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Barbier, M.</au><au>Gray, B. D.</au><au>Muirhead, K. A.</au><au>Ronot, X.</au><au>Boutonnat, J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis</atitle><jtitle>Cytometry. Part B, Clinical cytometry</jtitle><addtitle>Cytometry B Clin Cytom</addtitle><date>2004-05</date><risdate>2004</risdate><volume>59B</volume><issue>1</issue><spage>46</spage><epage>53</epage><pages>46-53</pages><issn>1552-4949</issn><eissn>1552-4957</eissn><abstract>Background
Proliferative status and multidrug resistance play a key role in determining cell response to chemotherapy. There is a need to develop multiple labeling method that allows simultaneous assessment of multidrug resistant (MDR) phenotype, proliferative status, apoptosis related changes in mitochondrial potential, in chemosensitive and chemoresistant tumor cell populations.
Methods
A three‐color labeling was performed using Hoechst 33342 (DNA), JC1 (mitochondrial potential), and a far red fluorescent membrane intercalating dye: PTIR271 (proliferation).
Results
Combined staining of DNA and mitochondrial potential allows identification of subpopulations expressing and MDR phenotype mediated by P‐glycoprotein (Pgp), and, in Pgp negative subpopulations, identification of apoptotic cells and evaluation of cell cycle status in viable cells. Addition of a far red fluorescent membrane intercalating dye, PTIR271, allows simultaneous monitoring of cell division status by dye dilution in both drug sensitive and drug resistant populations.
Conclusion
This triple labeling is an interesting method to study the proliferation status of drug sensitivity and drug resistance in viable tumor cells. © 2004 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>15108170</pmid><doi>10.1002/cyto.b.20004</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete; Wiley Online Library Free Content; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Antineoplastic Agents, Phytogenic - pharmacokinetics Antineoplastic Agents, Phytogenic - pharmacology apoptosis Apoptosis - drug effects Camptothecin - analogs & derivatives Camptothecin - pharmacokinetics Camptothecin - pharmacology Cell Division - drug effects Drug Resistance, Neoplasm Flow Cytometry - methods Fluorescent Dyes Humans K562 Cells MDR PKH dyes, PTIR271 Predictive Value of Tests proliferation monitoring Staining and Labeling - methods |
| title | A flow cytometric assay for simultaneous assessment of drug efflux, proliferation, and apoptosis |
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