磁性纳米四氧化三铁协同表柔比星逆转MCF-7/ADR细胞多药耐药研究  

Effect of magnetic nanoparticles of Fe_3O_4 and epirubicin on the reversal of multidrug resistance in human breast cancer MCF-7/ADR cells

作　　者：叶园园[1,2] 陈园园[1,2] 徐枫[1,2] 刘颖斌[1,2] 刘培峰[3] 韩宝三[1,2] 

机构地区：[1]上海交通大学医学院附属新华医院普外科,上海200092 [2]上海交通大学医学院胆道疾病研究所,上海200092 [3]上海交通大学医学院附属仁济医院上海市肿瘤研究所,上海200032

出　　处：《中华肿瘤防治杂志》2015年第18期1428-1433,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172078);上海交通大学医工交叉基金(YG2013MS16)

摘　　要：目的探讨磁性纳米四氧化三铁(MgNPs-Fe3O4)联合表柔比星(epirubicin,EPI)逆转人源乳腺癌细胞MCF-7/ADR耐药性的机制。方法将乳腺癌细胞株MCF-7/ADR分为对照组、MgNPs-Fe3O4组、EPI组和EPI+MgNPsFe3O4组,CCK8法检测MgNPs-Fe3O4和EPI对MCF-7/ADR细胞增殖的抑制作用;Hoechst33342法检测细胞凋亡;RT-PCR检测耐药基因ABCB1和ABCG2mRNA表达水平变化;蛋白质印迹法分析耐药蛋白P-gp、BCRP及其表达率。结果 CCK8法检测结果显示,EPI和EPI+MgNPs-Fe3O4对MCF-7/ADR细胞增殖抑制作用呈浓度依赖性,5μmol/L的EPI对MCF-7/ADR细胞增殖抑制率为(27.59±2.32)%,5μmol/L EPI+20μg/mL MgNPs-Fe3O4对MCF-7/ADR细胞增殖抑制率为(49.70±3.60)%,P=0.002。Hoechst33342荧光结果显示,EPI+MgNPs-Fe3O4组与其他3组相比,细胞凋亡率为(69.34±1.53)%,细胞凋亡的形态变化更加明显,高于对照组(3.33±0.58)%、MgNPs-Fe3O4组(4.33±0.58)%和EPI组(33.33±2.08)%。实时定量PCR结果显示,EPI+MgNPs-Fe3O4组ABCB1mRNA的相对表达量为0.347±0.012,低于对照组(1.001±0.072)、MgNPs-Fe3O4组(0.971±0.015)和EPI组(0.622±0.043);EPI+MgNPs-Fe3O4组的ABCG2mRNA的相对表达量为0.528±0.068,低于对照组(1.002±0.073)、MgNPs-Fe3O4组(0.949±0.026)和EPI组(0.785±0.083)。蛋白质印迹法结果显示,EPI+MgNPs-Fe3O4组的P-gp蛋白的相对表达量为0.363±0.025,低于对照组(0.957±0.031)、MgNPs-Fe3O4组(0.887±0.038)和EPI组(0.680±0.056);EPI+MgNPs-Fe3O4组BCRP蛋白的相对表达量为0.387±0.035,低于对照组(0.947±0.015)、MgNPs-Fe3O4组(0.933±0.040)和EPI组(0.633±0.050)。结论MgNPs-Fe3O4可能通过下调耐药基因ABCB1、ABCG2和耐药蛋白P-gp、BCRP的表达,逆转乳腺癌细胞对EPI的耐受性,增强化疗药物敏感度。OBJECTIVE To explore the mechanisms of magnetic nanoparticles of Fe3O4 on reversing the drug-resistance of epirubicin（EPI）in breast cancer MCF-7/ADR cell line.METHODS Human breast cancer cells MCF-7/ADR were divided into control group,MgNPs-Fe3O4 group,EPI group,and EPI＋MgNPs-Fe3O4 group.Inhibitory effect on the proliferation of MCF-7/ADR cells was evaluated by CCK8 method.Cell apoptosis was detected by Hoechst33342.The expression levels of ABCB1,and ABCG2 mRNA was detected by RT-PCR.The alteration of P-gp,and BCRP protein expression was detected by western blotting.RESULTS CCK8 assay showed that EPI and EPI＋ MgNPs-Fe3O4 decreased the proliferation of MCF-7/ADR cells in a dose-dependent manner,proliferation inhibition rate of 5μmol/L EPI on MCF-7/ADR cells was（27.59±2.32）%,5μmol/L EPI＋20μg/mL MgNPs-Fe3O4 on MCF-7/ADR cells was（49.70±3.60）%（P=0.002）.Hoechst33342 staining showed that cell apoptosis of EPI＋ MgNPs-Fe3O4 group was（69.34±1.53）%,higher than those in control group（3.33±0.58）%,MgNPs-Fe3O4group（4.33±0.58）%and EPI group（33.33±2.08）%.Real-time PCR results indicated that the expression level of ABCB1 mRNA of EPI＋ MgNPs-Fe3O4 group was 0.347±0.012,lower than those in control group（1.001±0.072）,MgNPs-Fe3O4group（0.971±0.015）and EPI group（0.622±0.043）.The relative expression level of ABCG2 mRNA of EPI＋ MgNPs-Fe3O4 group was 0.528±0.068,lower than those in control group（1.002±0.073）,MgNPs-Fe3O4group（0.949±0.026）and EPI group（0.785±0.083）.Western blot results showed that the expression level of P-gp protein of EPI＋ MgNPs-Fe3O4 group was 0.363±0.025,lower than those in control group（0.957±0.031）,MgNPs-Fe3O4group（0.887±0.038）,and EPI group（0.680±0.056）.The relative expression level of BCRP protein of EPI＋ MgNPs-Fe3O4 group was 0.387±0.035,lower than those in control group（0.947±0.015）,MgNPs-Fe3O4group（0.933±0.040）,and EPI group（0.633±0.050）.CONCLUSION MgNPs-Fe3O4 probably reverse epirubicin resista

关 键 词：磁性纳米四氧化三铁 表柔比星 乳腺肿瘤 耐药 逆转 

分 类 号：R737.9[医药卫生—肿瘤]