双硫仑螯合铜对耐紫杉醇乳腺癌细胞增殖、迁移、凋亡及自噬的影响  

Effect of DSF-Cu on the proliferation, migration, apoptosis and autophagy of paclitaxel-resistant breast cancer cells

作　　者：柴效科 周聪 宋爱琳[1] CHAI Xiaoke;ZHOU Cong;SONG Ailin(The Fifth Ward of General Surgery Department,The Second Hospital of Lanzhou University,Lanzhou,Gansu 730000,China)

机构地区：[1]兰州大学第二医院普外科5病区,甘肃兰州730000

出　　处：《安徽医药》2022年第12期2383-2389,共7页Anhui Medical and Pharmaceutical Journal

摘　　要：目的 探讨双硫仑螯合铜(DSF-Cu)对耐紫杉醇乳腺癌细胞增殖、迁移、凋亡及自噬的影响。方法 2018年1月至2019年10月,培养MCF-7人乳腺癌细胞和耐紫杉醇乳腺癌MCF-7/紫杉醇细胞,用定量紫杉醇持续诱导MCF-7/紫杉醇细胞维持耐药性。通过CCK-8法检测MCF-7/紫杉醇细胞耐药指数;通过CCK-8法检测MCF-7/紫杉醇细胞在不同浓度DSF-Cu(5、10、15、20、25μmol/L)干预不同时间(24 h和48 h)后的抑制率,并分析细胞抑制率与DSF-Cu浓度和作用时间的关系;通过划痕实验检测DSF-Cu对MCF-7/紫杉醇细胞迁移能力的影响;通过实时荧光定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹法检测MCF-7和MCF-7/紫杉醇细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和自噬效应蛋白Beclin-1基因和蛋白的表达,并在紫杉醇和DSF-Cu单药及联合干预MCF-7/紫杉醇细胞中后,检测Bcl-2、Bax和Beclin-1蛋白的表达;通过流式细胞计数技术检测不同浓度DSF-Cu(5、10、15、20μmol/L)干预MCF-7/紫杉醇细胞不同时间(12 h、24 h和48 h)后细胞凋亡情况。结果MCF-7和MCF-7/紫杉醇细胞在紫杉醇干预24 h后IC_(50)值分别为1 394.79μmol/L和321.42μmol/L,MCF-7/紫杉醇细胞耐药指数为4.34;在不同浓度DSF-Cu(5、10、15、20、25μmol/L)DSF-Cu干预MCF-7/紫杉醇细胞24 h后细胞抑制率分别为(27.42±3.38)%、(42.71±0.77)%、(57.52±7.10)%、(73.83±1.76)%、(85.55±2.25)%,48 h后细胞抑制率分别为(51.27±4.59)%、(57.34±5.94)%、(71.50±1.80)%、(81.54±4.36)%、(90.96±2.13)%,DSF-Cu以浓度与时间依赖性抑制乳腺癌MCF-7/紫杉醇细胞增殖(P<0.05);划痕实验证实DSF-Cu能抑制乳腺癌MCF-7/紫杉醇细胞迁移;qRT-PCR结果显示,与MCF-7细胞相比,MCF-7/紫杉醇细胞中Bcl-2和Beclin-1基因表达升高,Bax基因表达降低(P<0.05);蛋白质印迹法结果显示,与MCF-7细胞比较,MCF-7/紫杉醇细胞中Bcl-2和Beclin-1蛋白表达升高,Bax蛋白表达降低(P<0.05)。MCF-7/紫杉醇细胞在紫�Objective To investigate the effect of disulfiram chelated with Cu(DSF-Cu) on the proliferation,migration,apoptosis and autophagy of paclitaxel-resistant breast cancer cells.Methods From January 2018 to October 2019,MCF-7 human breast cancer cells and paclitaxel-resistant breast cancer MCF-7/paclitaxel cells were cultured,and MCF-7/paclitaxel cells were continuously induced to maintain drug resistance with quantitative paclitaxel.The drug resistance index of MCF-7/paclitaxel cells was detected by CCK-8 assay;the inhibition rate of MCF-7/paclitaxel cells after different concentrations of DSF-Cu(5,10,15,20,25 μmol/L) interventions for different times(24 h and 48 h) was detected by CCK-8 assay,and the relationship between cell inhibition rate and DSF-Cu concentration and duration of action was analyzed;the inhibition rate of DSF-Cu on the migration ability of MCF-7/paclitaxel cells was examined by scratch assay.Real-time quantitative polymerase chain reaction(qRT-PCR) and Western blotting were used to detect the expression of B-cell lymphoma-2(Bel-2),Bcl-2-associated X protein(Bax) and Beclin gene and protein in MCF-7 and MCF-7/paclitaxel cells after combined intervention in MCF-7/paclitaxel,and Bcl-2,Bax and Beclin-1 protein expression in MCF-7/paclitaxel cells after single and combined intervention with paclitaxel and DSF-Cu.Apoptosis was detected by flow cytometry after different concentrations of DSF-Cu(5,10,15 and 20 μmol/L) interventions on MCF-7/paclitaxel cells at different times(12 h,24 h and 48 h).Results The IC_(50)values of MCF-7 and MCF-7/paclitaxel cells were 1 394.79 μmol/L and 321.42 μmol/L,respectively.After 24 h of paclitaxel intervention,the MCF-7/paclitaxel cell resistance index was 4.34.The inhibition rates of MCF-7/paclitaxel cells were(27.42±3.38)%,(42.71±0.77)%,(57.52±7.10)%,(73.83±1.76)%,and(85.55±2.25)% after 24 h of intervention with different concentrations of DSF-Cu(5,10,15,20,25 μmol/L),and were(51.27±4.59)%,(57.34±5.94)%,(71.50±1.80)%,(81.54±4.36)%,and(90.96±2.13)% after

关 键 词：乳腺肿瘤 双硫仑 紫杉醇 抗药性 肿瘤 BCL-2相关X蛋白质 自噬相关蛋白质类 

分 类 号：R96[医药卫生—药理学]