三氧化二砷对恶性黑素瘤A_(375)细胞NF-κB、C-IAP2及caspase-3表达的影响

The effect of As_2O_3 on expressions of NF-κB,C-IAP2 and caspase-3 in melanoma A_(375) cell lines

目的：探讨三氧化二砷（arsenic trioxide,As2O3）在诱导人黑素瘤A375细胞凋亡中,对核因子-κB（NF-κB）、细胞凋亡抑制蛋白2（C-IAP2）以及半胱氨酸-天冬氨酸特异性蛋白酶-3（caspase-3）表达的影响。方法：Western blot法检测A375细胞中NF-κB p65核蛋白的表达和caspase-3蛋白的活化,半定量RT-PCR法检测C-IAP2mRNA的表达。结果：2.5、5.0、10.0μmol/LAs2O3作用于A375细胞24h,NF-κB p65核蛋白表达比值分别为（39.60±7.76）%、（10.17±0.90）%和（4.43±0.91）%;caspase-3蛋白比值分别为（10.03±2.06）%、（23.43±3.28）%和（35.70±4.61）%;C-IAP2mRNA表达的比值为（66.78±15.46）%、（30.16±5.52）%和（6.46±4.54）%,上述各指标与相应对照组比较及组内两两比较差异均有统计学意义（P〈0.05~P〈0.01）,随着As2O3作用浓度的增加,caspase-3激活型蛋白增多,NF-κB p65核蛋白和C-IAP2mRNA表达下降。结论：As2O3能抑制NF-κB、C-IAP2基因的表达,活化caspase-3蛋白表达。推测NF-κB-C-IAP2-caspase-3通路可能是As2O3诱导A375细胞凋亡的途径之一。

Objective：To explore the effect of As2O3 on expressions of NF-κB,C-IAP2 and caspase-3 in melanoma A375 cell line.Methods：The expression of NF-κB p65 nuclear protein and the activity of caspase-3 were examined by Western blot,the expression of C-IAP2 mRNA was examined by semi-quantitative RT-PCR in A375 cells.Results：Treated with 2.5,5.0 and 10.0 μmol/L As2O3 for 24 h,the expression ratio of NF-κB p65 nucleoprotein was（39.60±7.76）%,（10.17±0.90）% and（4.43±0.91）%,respectively,the expression ratio of cleaved caspase-3 was（10.03±2.06）%,（23.43±3.28）% and（35.70±4.61）%,respectively,the ratio of C-IAP2 mRNA was（66.78±15.46）%,（30.16±5.52）% and（6.46±4.54）%,respectively.There was significant difference between control group and experiment group（P〈0.05 to P〈0.01）（P〈0.01）.With the concentration of As2O3 increasing,caspase-3 was activated dramatically,NF-κB p65 and C-IAP2 mRNA decreased dramatically.Conclusions：The signal transduction pathway of NF-κB,C-IAP2 and caspase-3 may be associated the apoptosis of A375 cells induced by As2O3.