Role of Cyclin B1Cdc2 Up-Regulation in the Development of Mitotic Prometaphase Arrest in Human Breast Cancer Cells Treated with Nocodazole 英文参考文献.docVIP

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Role of Cyclin B1Cdc2 Up-Regulation in the Development of Mitotic Prometaphase Arrest in Human Breast Cancer Cells Treated with Nocodazole 英文参考文献.doc

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Role of Cyclin B1Cdc2 Up-Regulation in the Development of Mitotic Prometaphase Arrest in Human Breast Cancer Cells Treated with Nocodazole 英文参考文献

RoleofCyclinB1/Cdc2Up-Regulationinthe DevelopmentofMitoticPrometaphaseArrestinHuman BreastCancerCellsTreatedwithNocodazole HyeJoungChoi,MasayukiFukui,BaoTingZhu* DepartmentofPharmacology,ToxicologyandTherapeutics,SchoolofMedicine,UniversityofKansasMedicalCenter,KansasCity,Kansas,UnitedStatesofAmerica Abstract Background:Duringanormalcellcycle,thetransitionfromG2phasetomitoticphaseistriggeredbytheactivationofthe cyclin B1-dependent Cdc2 kinase. Here we report our finding that treatment of MCF-7 human breast cancer cells with nocodazole, a prototypic microtubule inhibitor, results in strong up-regulation of cyclin B1 and Cdc2 levels, and their increasesarerequiredforthedevelopmentofmitoticprometaphasearrestandcharacteristicphenotypes. Methodology/PrincipalFindings:Itwasobservedthattherewasatime-dependentearlyincreaseincyclinB1andCdc2 proteinlevels(peakingbetween12and24hposttreatment),andtheirlevelsstartedtodeclineaftertheinitialincrease.This earlyup-regulationofcyclinB1andCdc2closelymatchedintimingthenocodazole-inducedmitoticprometaphasearrest. Selective knockdown of cyclin B1or Cdc2 each abrogated nocodazole-induced accumulation of prometaphase cells. The nocodazole-induced prometaphase arrest was also abrogated by pre-treatment of cells with roscovitine, an inhibitor of cyclin-dependent kinases, or with cycloheximide, a protein synthesis inhibitor that was found to suppress cyclin B1 and Cdc2up-regulation.Inaddition,wefoundthatMAD2knockdownabrogatednocodazole-inducedaccumulationofcyclinB1 andCdc2proteins,whichwasaccompaniedbyanattenuationofnocodazole-inducedprometaphasearrest. Conclusions/Significance: These observations demonstrate that the strong early up-regulation of cyclin B1 and Cdc2 contributes critically to the rapid and selective accumulation of prometaphase-arrested cells, a phenomenon associated withexposuretomicrotubuleinhibitors. Citation:ChoiHJ,FukuiM,ZhuBT(2011)RoleofCyclinB1/Cdc2Up-RegulationintheDevelopmentofMitoticPrometaphaseArrestinHumanBreastC