咖啡因对p53缺失型人肺腺癌细胞h1299的放射增敏作用与机制分析

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1、中文摘要咖啡因对P53缺失型人肺腺癌细胞H1299的放射增敏作用及机制研究期阻滞的过程相一致。结论(1)咖啡因能抑fl刘p53缺失型人肺腺癌H1299细胞的生长；(2)咖啡因能增加p53缺失型人肺腺癌H1299细胞的放射敏感性；(3)咖啡因可以解除X射线诱导的G2期阻滞，抑制照射所致的DNA损伤的修复，促进照射所致的细胞凋亡；(4)咖啡因能降低照射后H1299细胞Cdc2mRNA的表达，表明咖啡因是通过改变细胞周期调控的信号因子来发挥放射增敏作用的。关键词人肺腺癌H1299细胞系；咖啡因；细胞凋亡；放射增敏作者：庞琴霞指导教师：高耀明张军宁IIaden

2、ocarcinomaH1299celllinebycaffeine，aswellastopreliminarilyinvestigateitsmechanism．MethodsHumanlungH1299celllinewimp53-deficientwasusedinthisstudy．(1)DetectingdifferentconcentrationsofcaffeineonH1299cellsgrowthinhibitionbytrypanblueexclusiontestandthesurvivalrateofHl299cellsaftert

3、heeffectofcaffeineatdifferenttimepointsbyMTTassay．(2)TheeffectofcaffeineonradiosensitivityWasdeterminedbyclonogenicassayandWasquantifiedbycalculatingthesensffiveenhancementratio(SER)．(3)FlowcytometryWasusedtoobservethecell-cycleeffectsofirradiationandcaffeineontheabrogationofrad

4、iation-inducedG2phasearrestinH1299cells．(4)TheeffectofcaffeineonapoptosisinirradiatedH1299cellsWasdeterminedbyflowcytometry．(5)Real—timequantitativePCRwasusedtoobservetheeffectofirradiationandcaffeineonCdc2mRNAexpressioninH1299cells．Results(1)Caffeinesignificantlyinhibitthegrowt

5、hofH1299cellsinaconcentration-·dependentandtime··dependentmanner．(2)CaffeinesignificantlyreducedH1299cellscloningefficiencyafterradiation．1mmol／Land2mmol／Lcaffeineresultedinaradiosensitization谢tllSERof1．20and2．07inH1299cell．(3)IrradiationresultedinG2phasearrestinH1299cellsinados

6、e·dependentmanner．At24hafterirradiationof0Gy，2Gy,4Gyand6Gy,theproportionofH1299cellsinG2phaseis13．800％，20．026％，26．929％and49．163％．(4)Radiation-inducedG2phasearrestWasabrogatedbycaffeineinaconcentration·dependentmanner．Caffeineintheconcentrationsoflmmol／L，2mmol／L，III、析mthearrestof

7、G2phaseafterirradiationinH1299cells．Caffeinecouldsurpresstheexpressionofradiation-inducedCdc2mRNA，whichwasinaccordancewiththeabrogationofradiation—inducedG2phasearrest．Conclusions(1)Caffeineisabletoinhibitp53一deficienthumanlungadenocarcinomaH1299celllinegrowth．(2)Caffeineisablet

8、opotentiatetheradiosensitivityofp53一deficienthu