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目的探讨Caspase8表达对肿瘤坏死因子相关凋亡诱导配体(tumornecrosisfactorrelatedapop-tosisinducingligand,TRAIL)诱导神经母细胞瘤(neuroblastoma,NB)细胞株SH-SY5Y(SY5Y)细胞凋亡的影响及其发生机制。方法应用RT-PCR方法和免疫组化法检测IFNγ作用前后SY5Y细胞Caspase8的表达;应用四甲基偶氮唑蓝(MTT)比色法及流式细胞仪(FCM)检测IFNγ、TRAIL、IFNγ+TRAIL及IFNγ+Cas-pase8抑制剂+TRAIL对SY5Y细胞生长及凋亡的影响。结果SY5Y细胞不表达Caspase8,IFNγ作用48h后的SY5Y细胞Caspase8表达明显增加;SY5Y细胞对TRAIL不敏感,经IFNγ诱导表达Caspase8的SY5Y细胞对TRAIL敏感,且与TRAIL浓度有关,Caspase8抑制剂可明显降低TRAIL对表达Caspase8的SY5Y细胞的杀伤作用。结论IFNγ通过诱导Caspase8表达而逆转SY5Y细胞对TRAIL诱导凋亡的耐受。
Objective To investigate the effect of Caspase8 expression on the apoptosis of neuroblastoma (NB-SY5Y) cells induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL) and its mechanism. Methods The expression of Caspase8 in SY5Y cells before and after IFNγ treatment was detected by RT-PCR and immunohistochemistry. The expressions of IFNγ, TRAIL and IFNγ + were detected by MTT assay and flow cytometry (FCM) Effect of TRAIL and IFNγ + Cas-pase8 inhibitor + TRAIL on the Growth and Apoptosis of SY5Y Cells. Results SY5Y cells did not express Caspase8, but Caspase8 expression was significantly increased in SY5Y cells treated with IFNγ for 48 hours. SY5Y cells were not sensitive to TRAIL. SY5Y cells expressing Caspase8 induced by IFNγ were sensitive to TRAIL and related to TRAIL concentration. Caspase8 inhibitor was significantly decreased The killing effect of TRAIL on SY5Y cells expressing Caspase8. Conclusion IFNγ reversed the tolerance of SY5Y cells to TRAIL-induced apoptosis by inducing Caspase8 expression.