CIK逆转耐顺铂人肺腺癌细胞对顺铂耐药与ERCC1的相关性-论文.pdf

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1、第23071卷4年第6月3期遵义医学院学报V01．37No．3JournalofZunyiMedicalUniversityJun．2014CIK逆转耐顺铂人肺腺癌细胞对顺铂耐药与ERCC1的相关性田昕，李宁，李冉(遵义医学院附属肿瘤医院胸部肿瘤科，贵州遵义563099)[摘要]目的观察脐血来源的CIK细胞对A549／DDP细胞的耐药逆转作用。并探讨CIK逆转耐药机制是否与核苷酸切除修复交叉互补基因1(excisionrepaircrosscomplementationgroup1，ERCC1)的

2、表达存在相关性。方法采集脐血单个核细胞经细胞因子体外诱导获得CIK细胞(cytokineinducedkillerceils，CIK)，MIrr法检测顺铂(cisplatin，DDP)对A549细胞和A549／DDP细胞的体外杀伤作用，以及CIK细胞与A549／DDP细胞共培养前后对DDP敏感性的变化。Real—timeRT—PCR法检测A549细胞、A549／DDP细胞以及CIK干预后的A549／DDP细胞中ERCC1mRNA的表达情况。Westernblot法检测A549细胞、A549／DDP

3、细胞以及CIK干预后的A549／DDP细胞中ERCC1蛋白的表达情况。结果A549／DDP细胞对DDP耐药，耐药指数为2．391。CIK细胞能逆转A549／DDP细胞对DDP的耐药性，逆转倍数为3．967。Real—timeRT—PCR、West—ernblot测得A549／DDP细胞中ERCC1mRNA和蛋白表达量比A549细胞高(P<0．05)，CIK细胞共培养前后A549／DDP细胞中ERCC1mRNA和蛋白表达量无明显变化。结论CIK细胞能逆转A549／DDP细胞对DDP的耐药性，其逆转耐

4、药的机制与ERCC1的表达无明显相关性。[关键词]肺癌；A549细胞；顺铂；耐药逆转；CIK细胞[中图法分类号]R734．2[文献标志码]A[文章编号]1000—2715(2014)03-0338-05StudyonthecorrelationbetweenERCC1andthemerchanismsunderlyingcy·tokineinducedkillercells——reversedcisplatinresistanceinA549／DDPCellsTianXin，LNing，LiRan

5、(DepartmentofThoracic，TheAffiliatedHospitalofZunyiMedicalUniversity，ZunyiGuizhou563099，China)[Abstract]ObjectiveToinvestigatetheeffectsofcordblood—derivedcytokine—inducedkiller(CIK)cellsonreversingcisplatinresistanceintheDDP——resistanthumanlungadenoca

6、rcinomacelllineA549／DDPandobservewhethertherewascorrelationbetweentheseeffectsandexcisionrepaircrosscomplementationgroup1(ERCC1)expression．MethodsThesingleumbilicalcordbloodmononuclearcellwasinducedbycytokineinvitrotobe—comeCIKcells．MTFassaywasapplied

7、toevaluatethecytotoxicactivityofDDPagainstA549／DDPcellsandA549cellsinvitroandthesensitivityofA549／DDPcellsCIKtoDDPbeforeandaftertheco—culturewithCIK．ThemRNAandproteinexpressionsofERCC1inA549cells。A549／DDPcellsandA549／DDPco—culturedwithCIKcellsweredete

8、ctedbyreal—timeRT—PCRandwesternblotanalysis．ResultsTheA549／DDPcellswasresistanttoDDPwiththeresistanceindexof2．391．CIKcellsdecreaseddrugresistancetocisplatinwiththere．versalfoldof3．967．TheERCC1mRNAandproteinexprossionswerehigherinA549／DDPthanth