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时间:2019-03-06
《rna干扰抑制ercc1对非小细胞肺癌化疗敏感性的影响[1]》由会员上传分享,免费在线阅读,更多相关内容在教育资源-天天文库。
1、3232重庆医学2011年11月第40卷第32期·论著·*RNA干扰抑制ERCC1对非小细胞肺癌化疗敏感性的影响宋志雨,周航△(遵义医学院附属医院肿瘤二科,贵州遵义563000)摘要:目的探讨利用RNA干扰技术沉默切除修复交叉互补基因组1(ERCC1)表达对非小细胞肺癌耐药细胞株顺铂化疗敏感性的影响。方法设计并合成3段靶向人的ERCC1基因的小分子干扰RNA(siRNA),构建携带ERCC1-shRNA的重组质粒表达载体,采用脂质体Lipofectamine2000转染入人肺癌细胞株A549/DDP,荧光镜下观察并测定转染效率;应用逆转录聚合酶链反应(RT-PCR)检
2、测转染前后ERCC1mRNA的表达情况;应用四甲基偶氮唑蓝比色法(MTT)检测干扰ERCC1后A549/DDP细胞对顺铂敏感性的变化。结果转染针对ERCC1的siRNA后,转染组A549/DDP细胞内ERCC1mRNA表达均下降,转染后肺癌A549/DDP细胞对顺铂敏感性增加。结论利用RNA干扰技术能够筛选出高效的特异阻断ERCC1基因表达的siRNA;ERCC1基因表达下调能够增加肺癌A549/DDP细胞对顺铂的敏感性,部分逆转耐药。关键词:RNA干扰;癌,非小细胞肺癌;顺铂;切除修复交叉互补基因组1;A549/DDP细胞doi:10.3969/j.issn.167
3、1-8348.2011.32.004文献标识码:A文章编号:1671-8348(2011)32-3232-04TheeffectofRNAinterference-mediatedERCC1geneonthechemo-treatmentsensitivityofnon-small-celllungcancer*SongZhiyu,ZhouHang△(TheSecondDepartmentofOncology,AffiliatedHospitalofZunyiMedicalCollege,Zunyi,Guizhou563000,China)Abstract:Objec
4、tiveToinvestigatechangesofplatinum-basedchemotherapysensitivityofsilencingexcisionrepaircrosscom-plementation1(ERCC1)geneexpressionbyusingRNAinterferenceinnon-small-celllungcancer(NSCLC)drugresistancecelllines.MethodsThreesiRNAsequencestargetingERCC1geneweredesignedandsynthesized.Recomb
5、inantplasmidexpressionvectorwhichcarryingERCC1-shRNAwasconstructedandtransfectedintoA549/DDPcellswithLipofectamine2000.Transfectionefficiencywasmeasuredinthefluorescentmicroscope.TheexpressionofERCC1mRNAwasdetectedbyreversetranscription-poly-merasechainreaction(RT-PCR).Thechangeofcispla
6、tinsensitivityafterinterferencewastestbyMTTassay.ResultsAftertrans-fectionofERCC1-siRNA,theERCC1mRNAexpressionsinA549/DDPcellswereallreduced.ThesensitivitytocisplatinofA549/DDPcelllinewasincreasedaftertransfection.ThesensitivitytocisplatinofA549/DDPcelllinewasincreasedaftertransfection.
7、ConclusionHighlyeffectiveandspecificsiRNAtargetingERCC1genecanbesuccessfullyscreenedbyRNAinterferencetech-nique.SelectivesilencingofERCC1genepromotesthesensitivitytocisplatininA549/DDPcellandcanpartlyreversethecisplatinresistanceinhumancisplatinresistantlungadeno-carcinomacelll
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