雷帕霉素衍生物fim-aap23573体外抗肿瘤活性及其机制的分析-antitumor activity and mechanism of rapamycin derivative fim - aap 23573 in vitro.docxVIP

10-7mol/L的FIM-A未对大鼠骨肉瘤UMR-106细胞mTOR、p70s6k及4E-BP1基因表达产生影响(P＞0.05)。【结论】1、大鼠骨肉瘤UMR-106细胞中mTOR信号通路呈现过度表达。2、雷帕霉素衍生物FIM-A（AP23573）在体外能抑制大鼠骨肉瘤UMR-106细胞增殖，最适宜的浓度约为10-7mol/L。3、其机制可能为使细胞周期停滞于G1期及抑制mTOR信号通路上p-mTOR、p-p70s6k、p-4EBP1蛋白磷酸化水平。【关键词】FIM-A;AP23573；骨肉瘤；mTOR。TheantitumoractivityandmechanismofFIM-A（AP23573）,anovelrapamycinanalog,onosteosarcomacellinvitroAbstractObjectToevaluatetheeffectofFIM-A(AP23573),anovelrapamycinanalog,ontheproliferation,apoptosisandcellcycleofUMR-106cells,andinvestigateitseffectsandmechanismsonPI3K/Akt/mTORpathway.MethodsUMR-106cellsweretreatedwithincreasingdosesofFIM-A.CCK-8wasusedtodetectUMR-106cellproliferation.RT-PCRwasusedtodetectthedifferenceofexpressionsofmTOR,p70s6kand4E-BP1mRNAbetweenrat’sosteoblastandUMR-106cell.AnnexinV-FITC/PIdoublestainingandflowcytometrywasusedtodetectcellapoptosis,andcellcycledistribution.TheexpressionsofmTOR,p70s6kand4E-BP1mRNAandtheirproteinphosphorylationweredetectedbyRT-PCRandWesternblotting,respectively.ResultsUMR-106cellhadsignificantlyhigherexpressionsofmTOR,p70s6kand4E-BP1mRNAthanosteoblastswhichwereisolatedfromnewbornratsandcollected,culturedandevaluatedinvitro.FIM-Ahadadose-dependentinhibitiononUMR-106cellproliferation,withamaximuminhibitoryeffectat10-7mol/L.ExposuretoFIM-Ainducedadose-dependentG1phasecellcyclearrestwithamaximumeffectat10-7mol/L.ExposuretoFIM-Ainducedadose-dependentinhibitionontheexpressionofphosphorylatedmTOR,p70s6kand4E-BP1,withamaximuminhibitoryeffectat10-7mol/L.However,nosignificantdifferentwasfoundontheeffectofapoptosisandmRNAexpressions.Conclusions1、UMR-106cellhadoverexpressionsofmRNAinmTORpathway.2、FIM-AeffectivelyinhibitedthegrowthofUMR-106cellsinvitro,withamostappropriateconcentrationat10-7mol/L.3、ThemechanismsmightbeFIM-AcaninhibitthecellcyclefromG1phasetoSphaseandreducetheexpressionofphosphorylatedmTOR,p70s6kand4E-BP1.KeywordsFIM-A;AP23573；osteosarcoma;mTOR.前言骨肉瘤(Oseteosarcoma)是最常见的原发性恶性骨肿瘤，其恶性程度较高，易早期发生肺部转移，预后较差。20世纪70年代前，骨肉瘤的主要治疗方法是高位截肢或关节离断，但仍有高达80%的患者一年内出现远处转移、5年生存率仅为20%左右[1