Imaging in real-time with FRET the redox response of tumorigenic cells to glutathione perturbations in a microscale flow
Despite the potential benefits of selective redox-modulating strategies for cancer therapy, an efficacious methodology for testing therapies remains elusive because of the difficulty in measuring intracellular redox potentials over time. In this report, we have incorporated a new FRET-based biosenso...
Saved in:
Published in | Integrative biology (Cambridge) Vol. 3; no. 3; p. 208 |
---|---|
Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
01.03.2011
|
Subjects | |
Online Access | Get more information |
Cover
Loading…
Abstract | Despite the potential benefits of selective redox-modulating strategies for cancer therapy, an efficacious methodology for testing therapies remains elusive because of the difficulty in measuring intracellular redox potentials over time. In this report, we have incorporated a new FRET-based biosensor to follow in real time redox-sensitive processes in cells transformed to be tumorigenic and cultured in a microfluidic channel. A microfluidic network was used to control micro-scale flow near the cells and at the same time deliver drugs exogenously. Subsequently, the response of a redox homeostasis circuit was tested, namely reduced glutathione (GSH)/oxidized glutathione(GSSG), to diamide, a thiol oxidant, and two drugs used for cancer therapies: BSO (L-buthionine-[SR]-sulfoximine) and BCNU (carmustine). The main outcome from these experiments is a comparison of the temporal depletion and recovery of GSH in single living cells in real-time. These data demonstrate that mammalian cells are capable of restoring a reduced intracellular redox environment in minutes after an acute oxidative insult is removed. This recovery is significantly delayed by (i) the inhibition of GSH biosynthesis by BSO; (ii) the inactivation of glutathione reductase by BCNU; and (iii) in tumorigenic cells relative to an isogenic non-tumorigenic control cell line. |
---|---|
AbstractList | Despite the potential benefits of selective redox-modulating strategies for cancer therapy, an efficacious methodology for testing therapies remains elusive because of the difficulty in measuring intracellular redox potentials over time. In this report, we have incorporated a new FRET-based biosensor to follow in real time redox-sensitive processes in cells transformed to be tumorigenic and cultured in a microfluidic channel. A microfluidic network was used to control micro-scale flow near the cells and at the same time deliver drugs exogenously. Subsequently, the response of a redox homeostasis circuit was tested, namely reduced glutathione (GSH)/oxidized glutathione(GSSG), to diamide, a thiol oxidant, and two drugs used for cancer therapies: BSO (L-buthionine-[SR]-sulfoximine) and BCNU (carmustine). The main outcome from these experiments is a comparison of the temporal depletion and recovery of GSH in single living cells in real-time. These data demonstrate that mammalian cells are capable of restoring a reduced intracellular redox environment in minutes after an acute oxidative insult is removed. This recovery is significantly delayed by (i) the inhibition of GSH biosynthesis by BSO; (ii) the inactivation of glutathione reductase by BCNU; and (iii) in tumorigenic cells relative to an isogenic non-tumorigenic control cell line. |
Author | Tsvid, Gene Gaskins, H Rex Trump, Lisa Rund, Laurie A Schook, Lawrence B Kolossov, Vladimir L Henderson, Jerrod L Timp, Gregory Kenis, Paul J A Lin, Chunchen Henry, Jennifer Jo |
Author_xml | – sequence: 1 givenname: Chunchen surname: Lin fullname: Lin, Chunchen email: clin7@nd.edu organization: University of Notre Dame, 316 Stinson-Remick Hall, South Bend, IN 46556. clin7@nd.edu – sequence: 2 givenname: Vladimir L surname: Kolossov fullname: Kolossov, Vladimir L – sequence: 3 givenname: Gene surname: Tsvid fullname: Tsvid, Gene – sequence: 4 givenname: Lisa surname: Trump fullname: Trump, Lisa – sequence: 5 givenname: Jennifer Jo surname: Henry fullname: Henry, Jennifer Jo – sequence: 6 givenname: Jerrod L surname: Henderson fullname: Henderson, Jerrod L – sequence: 7 givenname: Laurie A surname: Rund fullname: Rund, Laurie A – sequence: 8 givenname: Paul J A surname: Kenis fullname: Kenis, Paul J A – sequence: 9 givenname: Lawrence B surname: Schook fullname: Schook, Lawrence B – sequence: 10 givenname: H Rex surname: Gaskins fullname: Gaskins, H Rex – sequence: 11 givenname: Gregory surname: Timp fullname: Timp, Gregory |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/21183971$$D View this record in MEDLINE/PubMed |
BookMark | eNo1kF1LwzAYhYMo7kNv_AGSP1BN0o80lzK2ORgIsvvxNn3TZaRNaVI2_70V9eYczrl4OJwFue18h4Q8cfbCWapeNbMVY0zy8w2Zc5nLRElWzsgihDNjRcZYdk9mgvMyVZLPyXXXQmO7htqODgguibZFerHxRDef6wONJ5z62l8nDb3vAlJvaBxbP9gGO6upRucCjZ42bowQT3YaRHsc4jhUEKcUfthAW6sHHzQ4pMb5ywO5M-ACPv75khw268PqPdl_bHert32iUyljooxC5IXJS0iNERmHWihVSQPCQJFmuda8yFVZcVRClalkyBD4dABoVSuxJM-_2H6sWqyP_WBbGL6O_w-Ib4q-XwA |
CitedBy_id | crossref_primary_10_1258_ebm_2011_011009 crossref_primary_10_1016_j_febslet_2013_02_003 crossref_primary_10_1016_j_bbrc_2013_09_011 crossref_primary_10_1016_j_cclet_2019_11_009 crossref_primary_10_1016_j_bios_2019_01_005 crossref_primary_10_1177_1535370214522179 crossref_primary_10_1016_j_biomaterials_2013_01_092 crossref_primary_10_1258_ebm_2012_011436 crossref_primary_10_1517_17460441_2011_586691 crossref_primary_10_1042_BST20130103 crossref_primary_10_1016_j_tibtech_2014_09_006 |
ContentType | Journal Article |
DBID | CGR CUY CVF ECM EIF NPM |
DOI | 10.1039/c0ib00071j |
DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed |
DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) |
DatabaseTitleList | MEDLINE |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: EIF name: MEDLINE url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search sourceTypes: Index Database |
DeliveryMethod | no_fulltext_linktorsrc |
Discipline | Biology |
EISSN | 1757-9708 |
ExternalDocumentID | 21183971 |
Genre | Research Support, U.S. Gov't, Non-P.H.S Journal Article Research Support, N.I.H., Extramural |
GrantInformation_xml | – fundername: NIBIB NIH HHS grantid: R21 EB004513 – fundername: NCI NIH HHS grantid: R33 CA137719 – fundername: NIBIB NIH HHS grantid: EB004513 |
GroupedDBID | --- -JG 0-7 0R~ 4.4 48X 53G 5WD 705 7~J AAEMU AAHBH AAIMJ AAIWI AAJAE AAPQZ AAPXW AARHZ AAUAY AAUQX AAVAP AAWGC ABEMK ABGFH ABJNI ABMNT ABPTD ABWST ABXOH ABXVV ACGFS ACIWK ACLDK ACPRK ACUFI ADBBV ADGKP ADGZP ADIPN ADMRA ADQBN ADRTK ADSRN ADVEK AEFDR AELWJ AENEX AESAV AETBJ AFFZL AFGWE AFLYV AFRAH AFVBQ AGINJ AGQXC AGSTE AHGCF AHGXI AJEEA ALMA_UNASSIGNED_HOLDINGS ANBJS ANLMG ANUXI APEMP ASKNT ASPBG ATGXG AUDPV AUNWK AVWKF AZFZN BCRHZ BEYMZ BSQNT C6K CAG CGR COF CUY CVF DU5 EBS ECM EE0 EF- EIF EJD F5P FEDTE FHSFR FLUFQ FOEOM FQBLK GAUVT H13 HVGLF HZ~ H~N J3G J3H J3I KBUDW KOP KSI KSN L-8 ML0 NLBLG NOMLY NPM O-G O9- OBOKY ODMLO OJZSN OK1 OWPYF R56 RCNCU RNS ROX ROYLF RPMJG RRC RSCEA RUSNO SMJ YXANX |
ID | FETCH-LOGICAL-c377t-9f9ee16f58a3ff241ad299b7fa2fa6345cc16598b1e9298370e0ea1071ac9d92 |
IngestDate | Tue Oct 15 23:43:28 EDT 2024 |
IsDoiOpenAccess | false |
IsOpenAccess | true |
IsPeerReviewed | true |
IsScholarly | true |
Issue | 3 |
Language | English |
LinkModel | OpenURL |
MergedId | FETCHMERGED-LOGICAL-c377t-9f9ee16f58a3ff241ad299b7fa2fa6345cc16598b1e9298370e0ea1071ac9d92 |
OpenAccessLink | https://europepmc.org/articles/pmc3418872?pdf=render |
PMID | 21183971 |
ParticipantIDs | pubmed_primary_21183971 |
PublicationCentury | 2000 |
PublicationDate | 2011-03-00 |
PublicationDateYYYYMMDD | 2011-03-01 |
PublicationDate_xml | – month: 03 year: 2011 text: 2011-03-00 |
PublicationDecade | 2010 |
PublicationPlace | England |
PublicationPlace_xml | – name: England |
PublicationTitle | Integrative biology (Cambridge) |
PublicationTitleAlternate | Integr Biol (Camb) |
PublicationYear | 2011 |
References | 14715295 - Anal Biochem. 2004 Feb 1;325(1):144-50 15493909 - J Am Chem Soc. 2004 Oct 27;126(42):13616-7 10389592 - Blood Cells Mol Dis. 1999 Apr;25(2):103-9 8329979 - Cell Mol Biol (Noisy-le-grand). 1993 Jun;39(4):389-96 17538714 - Lab Chip. 2007 Jun;7(6):726-30 16402904 - Annu Rev Pharmacol Toxicol. 2006;46:215-34 10924858 - Free Radic Biol Med. 2000 May 1;28(9):1387-404 7651192 - Methods Enzymol. 1995;251:123-33 16669756 - Annu Rev Plant Biol. 2006;57:79-107 12727827 - Cancer Res. 2003 May 1;63(9):2109-17 10647931 - Cell. 2000 Jan 7;100(1):57-70 21606117 - Exp Biol Med (Maywood). 2011 Jun 1;236(6):681-91 18977455 - Mutat Res. 2009 Mar 31;674(1-2):36-44 16269193 - Trends Biotechnol. 2005 Dec;23(12):605-13 11038270 - Anal Biochem. 2000 Nov 1;286(1):35-7 11368918 - Free Radic Biol Med. 2001 Jun 1;30(11):1191-212 15138307 - Physiol Genomics. 2004 Jul 8;18(2):196-205 9278050 - Nature. 1997 Aug 28;388(6645):882-7 11566809 - Biophys J. 2001 Oct;81(4):2395-402 5353890 - Biochem Biophys Res Commun. 1969 Nov 6;37(4):593-6 8144521 - J Biol Chem. 1994 Apr 1;269(13):9397-400 7476363 - Methods Enzymol. 1995;252:283-92 18469822 - Nat Methods. 2008 Jun;5(6):553-9 19294303 - Lab Chip. 2009 Apr 7;9(7):925-34 11013257 - J Biol Chem. 2000 Dec 22;275(51):40180-6 14722062 - J Biol Chem. 2004 Mar 26;279(13):13044-53 14985369 - J Biol Chem. 2004 May 21;279(21):22284-93 10082535 - Mol Cell Biol. 1999 Apr;19(4):2690-8 10762593 - Front Biosci. 2000 Apr 1;5:D406-23 11795594 - IUBMB Life. 2001 Jul;52(1-2):55-9 7476373 - Methods Enzymol. 1995;252:38-53 19486941 - Free Radic Biol Med. 2009 Nov 1;47(9):1282-93 15958525 - Am J Physiol Cell Physiol. 2005 Nov;289(5):C1220-8 17335279 - J Am Chem Soc. 2007 Mar 28;129(12):3458-9 10944195 - Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):10032-7 3053703 - J Biol Chem. 1988 Nov 25;263(33):17205-8 20091013 - Lab Chip. 2010 Feb 7;10(3):388-91 6137189 - Annu Rev Biochem. 1983;52:711-60 16964292 - Oncogene. 2007 Feb 15;26(7):1038-45 19166318 - Biol Chem. 2009 Mar;390(3):191-214 10218110 - Adv Enzymol Relat Areas Mol Biol. 1999;73:209-67, xii 18222979 - Exp Biol Med (Maywood). 2008 Feb;233(2):238-48 15180957 - FASEB J. 2004 Aug;18(11):1246-8 8402615 - Cancer Res. 1993 Oct 1;53(19):4469-73 16677081 - Antioxid Redox Signal. 2006 Mar-Apr;8(3-4):354-61 11689426 - EMBO J. 2001 Nov 1;20(21):5853-62 14637055 - J Theor Biol. 2004 Jan 7;226(1):65-8 11516525 - Toxicology. 2001 Jun 21;163(2-3):153-62 |
References_xml | |
SSID | ssj0064004 |
Score | 2.01696 |
Snippet | Despite the potential benefits of selective redox-modulating strategies for cancer therapy, an efficacious methodology for testing therapies remains elusive... |
SourceID | pubmed |
SourceType | Index Database |
StartPage | 208 |
SubjectTerms | Animals Bacterial Proteins - chemistry Bacterial Proteins - genetics Biosensing Techniques - methods Buthionine Sulfoximine - pharmacology Carmustine - pharmacology Cell Line, Transformed Cell Tracking - methods CHO Cells Cricetinae Cricetulus Diamide - metabolism Diamide - pharmacology Fibroblasts - drug effects Fibroblasts - metabolism Fluorescence Resonance Energy Transfer - methods Glutathione - antagonists & inhibitors Glutathione - metabolism Glutathione Disulfide - metabolism Green Fluorescent Proteins - chemistry Green Fluorescent Proteins - genetics Kinetics Luminescent Proteins - chemistry Luminescent Proteins - genetics Microfluidic Analytical Techniques - methods Microscopy, Confocal Microscopy, Fluorescence - methods Oxidation-Reduction Oxidative Stress - drug effects Oxidative Stress - physiology Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Swine Transfection |
Title | Imaging in real-time with FRET the redox response of tumorigenic cells to glutathione perturbations in a microscale flow |
URI | https://www.ncbi.nlm.nih.gov/pubmed/21183971 |
Volume | 3 |
hasFullText | |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1Lb9QwELa2ICQuiPcb-cBtlZJsnj4i1Krl0QMKUm-V49iQarNZ7W5L4dfwU_nGTjamPARcrCiOI8vzefx5PJ5h7LkMwwqsIQ90GMogUWESVFhnMN21ELUBRhSd6L47yg4-JK-P0-PJ5JvntXS2qXbV11_eK_kfqeId5Eq3ZP9Bstuf4gWeIV-UkDDKv5LxYetyDNk7KXIeUKJ4Z1ndf79XOu9BXXcXKK0nrPMIOGspG5amzDdktrchHj6im-SH2IFzLvUK61A1epnLaUtue2uIU0_NvPvsM9rDPt6E9YLvIzrRyfBwF8wzNbxt-hN-rKWfxitob9AIA3VuXW7nsm7aZjXd2qTL9Xnj8lrr0QOgXAGFzqSwlr7hIvI8t3a1U7Z5mgciDwtfG8ce6GJfs7rPftL4YUwBU1XY06VT_yNIa9la2WObCybokr38ufZS9O2haoft5AVp0COyBrmVPiP9N4S8jcWLsRM2xLRreGm7YmlLeZPd6Pcb_KUDzy020Yvb7JrLQPrlDrvoIcSbBd9CiBOEOEGIA0LcQogPEOKd4R6EuIUQ33TcgxD_AUL0b8lHCHGC0F1W7u-Vrw6CPhlHoOI83wTCCK2jzKSFjI0B75M1mEyVGzkzMouTVKkoS0VRYZLPBIVU0qGW0AKRVKIWs3vsygIdeMB4oqJCpQq8VuhE4b9piqahAW-i4JbRQ3bfjdjJ0gVcORnG8tFvax6z6yPEnrCrBjNcPwVd3FTPrMy-AxwXbvY |
link.rule.ids | 786 |
linkProvider | National Library of Medicine |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Imaging+in+real-time+with+FRET+the+redox+response+of+tumorigenic+cells+to+glutathione+perturbations+in+a+microscale+flow&rft.jtitle=Integrative+biology+%28Cambridge%29&rft.au=Lin%2C+Chunchen&rft.au=Kolossov%2C+Vladimir+L&rft.au=Tsvid%2C+Gene&rft.au=Trump%2C+Lisa&rft.date=2011-03-01&rft.eissn=1757-9708&rft.volume=3&rft.issue=3&rft.spage=208&rft_id=info:doi/10.1039%2Fc0ib00071j&rft_id=info%3Apmid%2F21183971&rft_id=info%3Apmid%2F21183971&rft.externalDocID=21183971 |