顺铂诱导三阴性乳腺癌4T1耐药小鼠模型的建立 .pdf

顺铂诱导三阴性乳腺癌4T1耐药小鼠模型的建立

盛佳钰;陈红风

【摘要】ObjectiveToestablishacisplatin-resistant4T1mousemodelof

triplenegativebreastcancer.Meth-odsAdrugresistantmicemodelwas

establishedwithcisplatin(DDP)inductionandin-vivo/in-vitro

tumorigenicap-proach.ItsresistancecharacteristicswereidentifiedbyMTT

assay.Changesofdrugresistancegene(MDR1,BCRP,MMP7,GST-π)and

protein(P-gp,BCRP,MMP7)expression,andphosphorate-Aktandtotal-

Aktproteinexpressionwereevaluatedbyreal-timePCR,

immunohistochemistryandwesternblotmethod,respectively.Small

animalliveimagingtechnologywasappliedtodetecttumor

growth.ResultsResistancefold(RF)ofcisplatin-resistant4T1mousemodel

was12.84.TheexpressionofMDR1,BCRP,MMP7,GST-πmRNAandP-gp,

BCRP,MMP7proteinsintheresistantmicewerehigherthanthatinthe

non-resistantmice.Theresultofwesternblotshowedthatastatistically

higherexpressionofp-Aktinresistantmicethanthatinnon-resistantmice

atproteinlevels(P0.01).Nosignificantdifferenceoftumorgrowthrate

wasobservedbetweennon-resistantandresistantmice(P0.05).Given

samedoseofDDP,resistantmiceshowedlowersensitivitythannon-

resistantmicesignificantly(P0.01).ConclusionsWehavesuccessfully

establishedacis-platin-resistanttriplenegativebreastcancermodelin

mice,whichprovidesanewplatformforfurtherstudyonchemoresis-tant

reversalstrategyandindividualizedclinicaltreatmentofthisdisease.%目的

建立一种耐药性稳定的三阴性乳腺癌4T1耐药小鼠模型,为研究体内肿瘤耐药机制

和逆转药物的筛选奠定实验基础.方法采用顺铂(DDP)低剂量诱导及体内外交叉致

瘤结合的方法建立三阴性乳腺癌耐药小鼠模型;MTT法检测细胞耐药特性;实时荧光

定量PCR法分析耐药相关基因MDR1、BCRP、MMP7及GST-π表达差异;免疫

组化分析耐药相关蛋白P-gp、BCRP、MMP7表达差异;蛋白印迹法检测磷酸化

Akt(phosphorate-Akt,p-Akt)和总Akt(total-Akt,t-Akt)蛋白表达;小动物成像检

测观察肿瘤生长情况.结果MTT显示建立的三阴性乳腺癌耐药4T1小鼠模型的耐

药指数为12.84;耐药小鼠肿瘤组织中MDR1、BCRP、MMP7、GST-π基因

mRNA的表达