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-  2015 

硒结合蛋白1通过抑制超氧化物歧化酶增强奥沙利铂的细胞毒作用 Selenium-Binding Protein 1 Enhances Oxaliplatin-Induced Cytotoxicity Through Inhibiting Superoxide Dismutase

Keywords: DLD-1细胞,硒结合蛋白-1,超氧化物歧化酶

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Abstract:

目的:探讨人硒结合蛋白1(SBP1)在结直肠癌DLD-1细胞中的表达水平对超氧化物歧化酶(SOD)的影响及其引起的DLD-1细胞对奥沙利铂的敏感性变化,揭示SBP1的部分抗肿瘤作用机制。方法:MTS法检测不同浓度奥沙利铂处理SBP1siRNA或SOD1siRNA转染细胞后的细胞存活率,CuZn/Mn-SOD活性检测试剂盒检测SBP1siRNA转染对DLD-1细胞SOD酶活性的影响;实时荧光定量PCR检测下调SBP1后,SOD1和SOD2mRNA水平的变化;Western blot法检测下调SBP1或SOD1后DLD-1细胞内SBP1、SOD1及SOD2蛋白水平的变化;超氧化物阴离子荧光探针DHE染色法检测抗肿瘤药物引起的细胞内超氧化物阴离子的水平。结果:下调SBP1后,奥沙利铂对DLD-1细胞的毒性作用[半数毒性浓度IC50为(36.7±1.6)μmol/L]相比对照组[IC50为(17.5±0.8)μmol/L]明显减小(P<0.05);同时下调SBP1和SOD1后,奥沙利铂对DLD-1细胞的毒性作用[IC50为(16.8±1.0)μmol/L]相比对照组[IC50为(17.3±1.2)μmol/L]无明显变化(P>0.05);下调SBP1分别在转录水平、蛋白水平及酶活性水平上调了SOD,并减少了由奥沙利铂引起的DLD-1细胞内产生的超氧阴离子。结论:下调SBP1能上调SOD的水平,进而减小了奥沙利铂对DLD-1细胞的毒性作用,故SBP1能通过抑制SOD增强奥沙利铂对DLD-1细胞的毒性作用

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