【作者】 常志坤；

【导师】 戚晓东；

【作者基本信息】 首都医科大学 ， 外科学， 2008， 硕士

【摘要】 目的探讨腺病毒介导的p53基因（Ad-p53）对人乳腺癌阿霉素耐药细胞株MCF-7/Adr多药耐药的逆转作用及其对耐药相关蛋白P-gp、TOPOⅡ和GST-π表达的影响。方法以人乳腺癌MCF-7细胞及其阿霉素耐药株MCF-7/Adr为实验对象,通过腺病毒介导的野生型p53基因转染乳腺癌耐药细胞株MCF-7/Adr,cck-8法检测MCF-7/Adr细胞对阿霉素的耐药倍数及基因转染后的耐药逆转倍数;采用免疫荧光组织化学法及western blot检测Ad-p53作用前后MCF-7/Adr细胞内P-gp、TOPOⅡ和GST-π蛋白表达的变化。结果50 MOI Ad-p53能使阿霉素对MCF-7/Adr细胞IC₅₀由（4.54±0.91）μg/ml降到（0.26±0.11）μg/ml,逆转耐药倍数为18.1倍（P<0.01）;免疫荧光组织化学及western blot显示P-gp、GST-π蛋白表达量下降,TOPOⅡ未见明显变化。结论Ad-p53对乳腺癌耐药细胞株MCF-7/Adr的耐药性有明显的逆转作用,其机制与下调P-gp和GST-π表达有关。更多还原

【Abstract】 ObjectiveTo explore whether adenovirus mediated p53 gene （Ad-p53） could reverse multidrug resistance（MDR） of human breast cancer and its impacts on the expressions of P-gp、TOPOⅡand GST-π.MethodsIn this study, adriamycin-resistant human breast carcinoma cells （MCF-7/Adr） and its parental cells （MCF-7） were used to determine the effect of Ad-p53. Wt-p53 gene was transfected into MCF-7/Adr cell line through adenovirus-mediated. Cck-8 assay was adopted to evaluate the cytotoxicity of adriamycin. Immunofluorescence histochemical and western blot were performed to observe the expression of P-glycoprotein（P-gp）、TopoismeraseⅡ（TOPOⅡ） and GST-π.ResultsAfter transfection with 50 MOI Ad-p53, the 50% inhibitory concentration (IC₅₀) of adriamycin to MCF-7/Adr cells was decreased from （4.54±0.91）μg/ml to （0.26±0.11）μg/ml, and the chemosensitivity increased 18.1 times （P<0.01）. Immunofluorescence histochemical and western blot analysis showed that the expressions of P-gp and GST-πwere down-regulated, and there is no significant change of TOPOⅡexpression.ConclusionsAd-p53 could reverse multidrug resistance of MCF-7/Adr cells through the down-regulation of P-gp and GST-πexpressions.更多还原