Survivin抑制剂YM155对甲状腺癌B-CPAP细胞凋亡的影响及机制研究  

Survivin inhibitor YM155 induces apoptosis of thyroid carcinoma cell line BCPAP and potential mechanisms

作　　者：王德平[1] 关悦[1] 李思瓯[1] 王云枫[1] 郭长秀[1] 孙毓晗[1] 

机构地区：[1]牡丹江医学院红旗医院,牡丹江157011

出　　处：《中国免疫学杂志》2017年第7期1023-1028,共6页Chinese Journal of Immunology

摘　　要：目的:探讨人生存素(Survivin)抑制剂YM155{4,9-二氢-1-(2-甲氧基乙基)-2-甲基-4,9-二氧代-3-(2-吡嗪甲基)-1H-萘并[2,3-d]咪唑鎓溴化物}对甲状腺癌B-CPAP细胞活力和凋亡的影响以及凋亡相关酶半胱氨酸蛋白酶-3(Cysteinyl aspartate specific proteinase-3,Caspase-3),半胱氨酸蛋白酶-8(Cysteinyl aspartate specific proteinase-8,Caspase-8)和半胱氨酸蛋白酶-9(Cysteinyl aspartate specific proteinase-9,Caspase-9)表达的变化,探讨其诱导B-CPAP细胞凋亡的可能机制。方法:体外培养B-CPAP细胞,分别以0、0.5、1、2、4、8 nmol/L浓度的YM155进行处理24、48和72 h,采用CCK-8检测试剂盒检测YM155对B-CPAP细胞活力的抑制作用;B-CPAP细胞随机分为4组:分别以0、1、2 nmol/L YM155和5μmol/L顺铂(Cisplatin,阳性对照组)处理24 h。分别采用TUNEL染色和流式细胞仪Annexin V-FITC/PI法检测凋亡的情况;采用RT-PCR检测Survivin mRNA的表达;采用比色法和Western blot检测Survivin和Caspase-3、Caspase-8、Caspase-9的活性和表达。结果:与0 nmol/L组比较,YM155对B-CPAP细胞活力具有明显抑制作用,并诱导B-CPAP细胞凋亡(P<0.05或P<0.01);与阴性对照组比较,YM155显著降低B-CPAP细胞Survivin的表达,并上调Caspase-3、Caspase-8、Caspase-9的表达(P<0.05或P<0.01)。结论:YM155对B-CPAP细胞活力具有抑制作用,诱导其凋亡,其机制可能与上调Caspase-3、Caspase-8和Caspase-9表达有关。Objective： To investigate the effects of survivin inhibitor YM155 （ 1- （ 2-methoxyethyl ） -2-methyl-4,9-dioxo-3- （pyrazin-2-ylmethyl）-4,9-dihydro-1H-naphtho [ 2,3-d ] imidazolium bromide / on cell viability, apoptosis and Cysteinyl aspartate specific proteinase-3, Cysteinyl aspartate specific proteinase-8, Cysteinyl aspartate specific proteinase-9 of the thyroid carcinoma cell line B-CPAP in order to discuss mitochondrial mechanisms of apoptosis. Methods： B-CPAP cells were cultured in vitro and treated with YM155 at various concentrations（0,0. 5,1,2,4,8 nmol/L）for 24,48 and 72h. The cell viability of B-CPAP cells were measured by CCK-8 assay. B-CPAP cells were randomly divided into 4 groups：B-CPAP cells were treated with YM155 at various concentrations （0, 1,2 nmol/L）and 5 μmol/L Cisplatin（the positive control group）for 24 h. The effects of YM155 on B-CPAP cells apoptosis were evaluated by TUNEL and flow cytometry Annexin V-FITC/PI method. The expression level of Survivin and Caspase-3, Caspase-8 , Caspase-9 were detected by Western blot analysis. Results： Compared with the 0 nmol/L group, YM155 significantly inhibited the cell viability of B-CPAP cells and induced their apoptosis （P〈0. 05 or P〈0. 01 ）. Compared with the 0 nmol/L group, YM155 significantly reduced the expression level of Survivin and upregulated Caspase-3, Caspase-8 , Caspase-9 （ P〈0.05 or P〈0. 01 ）. Conclusion ： YM155 can inhibit the cell viability of B-CPAP cells and induce apoptosis, its possible mechanisms maybe related to upregulated expression level of Caspase-3, Caspase-8 and Caspase-9.

关 键 词：YM155 甲状腺癌 凋亡 半胱氨酸蛋白酶-3 半胱氨酸蛋白酶-8 半胱氨酸蛋白酶-9 

分 类 号：R737.9[医药卫生—肿瘤]