NSC23925, Identified in a High-Throughput Cell-Based Screen, Reverses Multidrug Resistance 英文参考文献.docVIP

NSC23925,IdentifiedinaHigh-ThroughputCell-Based Screen,ReversesMultidrugResistance ZhenfengDuan*,EdwinChoy,FrancisJ.Hornicek SarcomaBiologyLaboratory,CenterforSarcomaandConnectiveTissueOncology,MassachusettsGeneralHospital,Boston,Massachusetts,UnitedStatesofAmerica Abstract Background: Multidrug resistance (MDR) is amajor factor which contributes tothe failure of cancer chemotherapy, and numerouseffortshavebeenattemptedtoovercomeMDR.Todate,noneoftheseattemptshaveyieldedatolerableand effectivetherapytoreverseMDR;thus,identificationofnewagentswouldbeusefulbothclinicallyandscientifically. Methodology/PrincipalFindings:Toidentifysmallmoleculecompoundsthatcanreversechemoresistance,wedeveloped a96-wellplatehigh-throughputcell-basedscreeningassayinapaclitaxelresistantovariancancercellline.Coincubating cellswithasublethalconcentrationofpaclitaxelincombinationwitheachof2,000smallmoleculecompoundsfromthe NationalCancerInstituteDiversitySetLibrary,weidentifiedapreviouslyuncharacterizedmolecule,NSC23925,thatinhibits Pgp1andreversesMDR1(Pgp1)butdoesnotinhibitMRPorBCRP-mediatedMDR.ThecytotoxicactivityofNSC23925was furtherevaluatedusingapanelofcancercelllinesexpressingPgp1,MRP,andBCRP.Wefoundthatataconcentrationof .10mMNSC23925moderatelyinhibitstheproliferationofbothsensitiveandresistantcelllineswithalmostequalactivity, butitsinhibitoryeffectwasnotalteredbyco-incubationwiththePgp1inhibitor,verapamil,suggestingthatNSC23925itself isnotasubstrateofPgp1.Additionally,NSC23925increasestheintracellularaccumulationofPgp1substrates:calceinAM, Rhodamine-123, paclitaxel, mitoxantrone, and doxorubicin. Interestingly, we further observed that, although NSC23925 directly inhibits the function of Pgp1 in a dose-dependent manner without altering the total expression level of Pgp1, NSC23925actuallystimulatesATPaseactivityofPgp,aphenomenonseeninotherPgpinhibitors. Conclusions/Significance: The ability of NSC23925 to restore sensitivity to the cytotoxic effects of chemotherapy or to preventresista