中文摘要 目的:应用荧光原位杂交技术检测人大肠癌细胞照射后2号染色体易位 畸变并分析其与照射后细胞存活率的相关性,探讨荧光原位杂交技术预测 人大肠癌细胞内在放射敏感性的可行性。 方法:体外实验部分:培养2种不同放射敏感性的人大肠癌细胞系Lov。 和SW48O,利用集落形成实验测得O、2、4、6GyX一线照射后的细胞存活 率,同时采用荧光原位杂交技术和2号染色体涂染探针检测O、2、4、6Gy X一线照射后24小时细胞2号染色体的诱导易位畸变量,分析染色体诱导 易位畸变量与细胞存活率二者的相关性。体内实验部分:将上述2种细胞 接种于裸鼠背部皮下,制成动物模型每组各12只,移植肿瘤最大径长至 0 . 4一0.6cm时予以10GyX一线局部照射。每组6只于照射后24小时取出 肿瘤,制成细胞悬液,短期培养后用荧光原位杂交技术检测并比较2种肿 瘤细胞的2号染色体诱导易位畸变量。另外6只隔日测量肿瘤最大径,绘 制肿瘤生长曲线,比较放疗对2种肿瘤生长的抑制差异。 结果:2种大肠癌细胞的存活曲线显示,Lov。细胞的放射敏感性高于 SW48O细胞。2种细胞照射后24小时的2号染色体诱导易位畸变量均随照 射剂量增大而增加。在同一剂量点,放射敏感性高的Lov。细胞的染色体 诱导畸变量显著大于放射敏感性低的Sw48O细胞(P<0.05)。对2种细 胞的实验数据行直线相关分析显示:2号染色体诱导易位畸变量与细胞存 活率显著相关(r=一0.89,P<0.05)。2种移植肿瘤照射后的生长曲线也荧光原位杂交技术预测人大肠癌细胞放射敏感性的可行性研究中文摘要 表明,放疗使Lov。移植瘤的增长明显延缓,Lov。细胞的放射敏感性高于 SW480细胞。X一线照射后肿瘤荧光原位杂交技术检测结果显示:Lov。细胞 的2号染色体诱导易位畸变量显著大于SW48O细胞(P<0.05)。 结论:荧光原位杂交技术能快捷、可靠地预测人大肠癌细胞的放射敏感 性,从而为临床大肠癌病人放疗的选择提供重要参考依据。 关键词荧光原位杂交预测大肠癌细胞放射敏感性 作者:李国强 指导教师:邢春根荧光原位杂交技术预测人大肠癌细胞放射敏感性的可行性研究英文摘要 TheFeasibilityStudyonPredietingtheRadiosensitivity ofHumanColoreetalCancerCellbyFluoreseeneeInSitu Hybridization ABSTRACT Objeetive:tostudytheeorrelationbetweenthenumberofradiation一indueed translocationsofchromosome2andthecellsurvivalfraetionafterradiation usingfluoresceneeinsituhybridization,anddiseussthefeasibilitytoPrediet theradiosensitivityofhumaneolorectaleaneercellusingFISH. Methods:Invitrostudy:TwohumaneolorectalcancercelllinesLovoand SW48OwereeultivatedandtheeellsurvivalfraetionafterradiationbyX一ray witho,2,4,6GywasdeteetedbymeansofelonogenieassayresPeetively. Theeellsurvivalcurvewasdrawn.Atthemeantimethenumberof radiatio一indueed一transloeationsofchromsome2wasobservedusingFISHand ehromosome2PaintingProbeatthetwenty一fourthhourafterradiation.Then theeorrelationbetweenthenumberofehromosomeaberrationsandtheeell survivalfractionwasobserved.Invivostudy:ThecellLovoandSW480were transPlantedintosubeutaneoustissueofnudemiee,and12animalmodels wereobtainedresPeetively.TheimPlantedtumorswereirradiatedwith10Gy whentheirdiameterswere4一6mm.imPlantedtumorsof6mieewerereseeted atthetwenty-fourihhourafterradiation,madeintosusPentionliquid,andeultivatedinvltroinshorttime.Thenumberofehromosomaltranslocationsof the2imPlantedtumorswaseomParedusingFISHandehromosome2Painting Probe.Totherest6miceeaehgrouP,thediameterofimPlantedtumorswas measuredevery2days,andthenthegroweurvewasdrawnthroughwhiehthe radiosensitivityofthe2eelllineswaseomPared. Results:Theeellsurvivaleurveshowedthattheradiosensitivityoftheeell 111 芝沪荧光原位杂交技术预测人大肠癌细胞放射敏感性的可行性研究英文摘要 lineLovowashigherthanSW480.Adose一dePendentinereasein radiation一indueedchromosomeaberrationswasobservedinthe2celllines. Onthesamedose,thenumberofehromosome2aberrationinLovoeellwas higherthanSW480signifieantly,ByintegratingtheexPerimentaldataoftheZ eelllines,Agoodeorrelation(厂一0.89,P<0.05)vvasfoundbetweeneell survivalandthenumberofradiation一indueedtransloeationsofchromosome2. ThegroweurveofimPlantedtumorafterradiationshowedthatthesensitivity ofLovoeellwashigherthanSW48O.TheresultofFISHdeteetionto imPlantedtumorreseetedindicatedthenumberofehromosome2aberrationof LovoeellwashigherthanSW480significantly(P<0.05). Conelusions:FluoresceneeInSituHybridizationeanbeusedtoPredictthe intrinsicradiosensitivityofhumaneoloreetaleaneereelleonvenientlyand reliably.ItProvidesimPortantrefereneesfortheseleetionofradiotheraPy inthePatientssufferingeoloreetaleaneereliniealy. KeyWords:eolorectaleaneerradiosensitivityPredietfluoreseeneein situhybridization Writtenby11guoqiang SuPervisedbyxingchungen 七目录 引言................................................1 体外实验部分............................................3 材料与方法.........................、..........3 结果...............................................11 讨论...............................................15 结论...............................................20