耐5-氟尿嘧啶的人结肠癌HT-29／5-FU细胞株的建立及其生物学特性.pdf

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1、328细胞与分子免疫学杂志(ChinJCellMolIminuno1)2015，31(3·论著·文章编号：1007—8738(2015)03—0328—05耐5-氟尿嘧啶的人结肠癌HT-29／5-FU细胞株的建立及其生物学特性邱婷婷，项晓军，雷婉，郝超，张凌，冯苗，余锋，黎军和，熊建萍(南昌大学第一附属医院肿瘤科，南昌大学泌尿外科研究所，江西南昌330006)[摘要]目的建立耐5．氟尿嘧啶人结肠癌细胞株(HT-29／5一FU)并观察其生物学特性。方法采用5·Fu持续接触浓度递增法诱导；观察细胞形态变化和克隆形成率；MTr法测定生长曲线和药物敏感性；Westernb

2、lot法分析胸苷酸合成酶(TS)、二氢嘧啶脱氢酶(DPD)的表达情况；流式细胞术检测细胞周期和凋亡。结果HT-29／5一FU的耐药指数为3．59；与HT-29细胞相比，HT．29／5．FU细胞形态发生改变、生长缓慢、克隆形成率降低，Ts和DPD高表达；s期细胞比例增多，更能耐受5一Fu诱导的细胞凋亡。结论HT-29／5一FU细胞株生物学性状的改变可能是5-Fu化疗耐药机制的重要组成部分，有助于阐明肿瘤潜在耐药机制和逆转肿瘤耐药。[关键词]结肠肿瘤；HT-29细胞；氟尿嘧啶；抗药性；细胞培养[中图分类号]R735．35，R392．5，Q279，R965[文献标志码]

3、AEstablishmentandbiologicalcharacterizationof5-fluorouracil·resistanthumancoloncancerHT-29／5．FUcelllineQIUTingting，XIANGXiaojun，LEIWan，HAOChao，ZHANGLing，FENGMiao，YUFeng’，LIJunhe，XIONGJianpingDepartmentofOncology，FirstAfiliatedHospital，NanchangUniversity；InstituteofUrology，NanchangUniv

4、ersity，Nanehang330006，ChinalAbstractlobjectiveToestablish5-fluorouraciI(5-Fu)·resistanthumancoloncancerHT-29cellline(HT-29／5~FU)vitroandobserveitsbiologicalproperties．MethodsTheHT-29／5-FUcelllinewasestablishedbycontinuouslyexposingtheHT-29celstoascendingdosesof5-FU．Themorphologyandcol

5、onyformationrateweredetected．ThegrowthcurveandthechemosensitivityofHT一29／5-FUcelIIineweredeterminedbVM1_rassay．Westernblottingwasusedtoanalyzetheexpressionsofthymidylatesynthase(TS)anddihydropyrimidinedehydrogenase(DPD)．Cellcycleandapoptosisweremeasuredbyflowcytometry．ResultsTheresist

6、anceofHT．29／5-FUcellsto5一FUwas3．59-foldgreaterthanthatofHT-29cells．ThemorphologyofHT-29／5-FUcellsdiferedfromthalOfHT-29cells．ComparedwithHT-29cells．HT一29／5-FUcellsshowedremarkablereductionofcellproliferationandcolonyformation．higherexpressionsofTSandDPD，higherpercentageofcellsintheSph

7、ase，andstrongerabilityofresistancetoapoptosisinducedby5一FU．ConclusionThebiologicalcharactersofHT-29／5-FUcelllinemayplayanimportantrolein5-FU-resistantmechanisms．whichmayalsocontributetoelucidatethepotentialmechanismsoftumordrugresistanceaswellasresistancereversa1．1Keywords]colonicneop

8、lasms