【摘要】 目的:研究四硫化四砷在抗急性早幼粒细胞白血病中诱导细胞凋亡的分子机制。方法:应用cDNA微矩阵技术检测四硫化四砷作用前后NB4细胞基因表达图谱。筛选出有差异表达的与细胞凋亡相关蛋白类基因,合成相应的引物,用RTPCR方法检测口服四硫化四砷的急性早幼粒细胞白血病(APL)患者缓解前后外周血细胞凋亡相关蛋白类基因的表达,分析四硫化四砷诱导早幼粒细胞凋亡的途径。结果:通过基因微矩阵技术分析发现,四硫化四砷作用NB4细胞有差异表达的细胞凋亡相关蛋白类基因包括编码凋亡蛋白酶活化因子基因(Apaf1),比值为2.910;编码细胞凋亡相关蛋白PNAS2基因,比值为0.420。RTPCR检测口服四硫化四砷APL患者缓解前后外周血Apaf1比值2.33,caspase9比值3.21,PNAS2比值0.99。结论:四硫化四砷对NB4细胞的细胞凋亡效应主要涉及两个基因表达的改变:细胞凋亡蛋白酶活化因子(Apaf1)基因的表达上调和细胞凋亡相关蛋白PNAS2基因表达的下调。APL患者PNAS2基因表达没有明显的改变,可能与使用其他药物的影响有关。Apaf1和caspase9表达相应的增高提示,四硫化四砷诱导APL细胞凋亡可能是通过以线粒体介导的凋亡通路为主的活化途径,而不是通过死亡受体途径。更多还原

【Abstract】 Objective: To investigate the mechanism of tetra-arsenic tetra-sulfide （As₄S₄） in inducing apoptosis of acute promyelocytic leukemia （APL） cells. Methods: The gene expression patterns in NB4 cells pre- and post-treatment with As₄S₄ were analyzed by cDNA microarray, and differentially expressed genes related with apoptosis were identified. The mRNA expression levels of these apoptosis related genes in the peripheral blood of APL patients treated with As₄S₄ pre-and post-remission were examined by RT-PCR. Results: Among the differentially expressed genes in NB4 cells pre- and post-treatment with As₄S₄, two genes were related with cellular apoptosis, which were Apaf-1 and PNAS-2. Apaf1 ratio between pre and post- As₄S₄ in NB4 cells was 2.910, PNAS-2 ratio was 0.420. RT-PCR results showed that the expression ratios of Apaf-1, caspase-9 and PNAS-2 in APL patients pre- and post-remission were 2.31 and 3.21, 0.99 respectively. Conclusion: As₄S₄ induced cellular apoptosis in NB4 cells involves the expression changes of the two genes: the up-regulation of Apaf1 and down-regulation of PNAS-2. The increased expressions of Apaf1 and caspase-9 indicate that As₄S₄ induced apoptosis in APL cells is through mitochondrial pathway, but not the death-receptor pathway. The role played by PNAS-2 in cellular apoptosis needs to be clarified.更多还原