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ID:53740928
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时间:2020-04-21
《MiR-192靶向负调控Bim表达诱导肺癌顺铂耐药-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、·384·扣国肺癌杂志2014年5月第17卷第5期ChinJLuGg!三!垫!!!:!!!:DOI:10.3779/j.issn.1009-3419.2014.05.04基础研究·MiR一192靶向负调控Bim表达诱导肺癌顺铂耐药张芳李洋吴蘅齐康尤嘉琮李雪冰祖玲玲潘振华王玉丽李永文李颖王珉沈旺周清华【摘要】背景与目的顺铂是非小细胞肺癌化疗的一线药物,但获得性耐药限制了其疗效的发挥。本研究的目的是筛选鉴定与肺癌顺铂耐药相关的microRNAs,探讨其与肺癌Jl~f]耐药的影响及分子机制。方法应用miRNA芯片及RT-PCR~选鉴定A549与A549/DDPIr~癌
2、细胞问的差异表达miRNAs,将差异表达miR-192转染A549和A549/DDP细胞株,CCK-8检测m192对半数抑制浓度(halfinhibitionconcentration,IC。)的影响,流式细胞术检测细胞凋亡,生物软件预测及双荧光素酶报告基因法寻找miR-192的靶基因,RT-PCR~Westernblot检测靶基因表达水平在转染前后的变化。结果MiR-192在A549/DDP中显著高表达,表达量是A549细胞表达量的37.59~0.35倍。在低表达miR-192的A549细胞中过表达miR.192,细胞对顺铂的Ic。显著增高,顺铂引起的细胞凋亡
3、率显著降低;反之,在高表达miRo192的A549/DDP中抑~miR-192的表达,顺铂的Ic显著降低,顺铂引起的细胞凋亡率显著增加。miR-192可靶向作用促凋亡基因Bim的3"UTR,并在转录后水平负向调控Bim的表达。结论MiR一192通过靶向负调控促凋亡基因Bim表达,诱导肺腺癌细胞株A549产生顺铂耐药,并减少顺铂引起的细胞凋亡。【关键词】肺肿瘤;mlKNA~t>。片;RT.PCR;miR-192;BimMiR-192ConfersCisplatinResistancebyTargetingBiminLungCancerFangZHANG,YangL
4、I,HengwUJKang03,JiacongYOUJXuebingLI,Linglingzu,ZhenhuaPAN,YuliWANG,YongwenLI,YingLI,MinWANG,WangSHEN,OjnghuaZHOUTianjinKeyLaboratoryofLungCancerMetastasisandTumorMc,.0e"fr0enTianjinLungCancerInstitute,TianjinMedicalUniversityGeneralHospital,Tianjin300052,ChinaZHANGandLIcontributedeq
5、uallytothiswork.Correspongdingauthor:QinghuaZHOU,E—maihzhouqh135@j63.com【Abstract】BackgroundandobjectiveCisplatinisthefirst—linedrugforthechemotherapyofnon—smallcelllungcancer(NSCLC)Jbuttheacquiredchemoresistancerestrictedtheefectofitstreatment.TheaimofthisstudyistovalidatethemiRNAsr
6、elatedtotheCisplatinresistanceinlungcancerandelucidatethemolecularmechanisms.MethodsWeperformedmiRNAmicroarrayandRT-PCRtoobtaintheaberrantdiferentialexpressedmiRNAsbetweenA549anditspairedCisplatin-resistantcelllineA549/DDPcells,andthenweinvestigatedthebiologicalfunctionsofmiR-192Jwhi
7、chistheaberrantdiferen-tialexpressedmiRNA.AftertransfectionofthemiR-192intoA549cellsJwemeasuredthehalfinhibitionconcentration(IC50),cellapoptosisofthetrasfectantcellsJandthenweusedbiologicalsoftwaresanddual—luciferasereportassaytoexplorethetargetgeneofthemiR-192,whichwasfurthervalida
8、tedbyRT-PCRa
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