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目的探讨IFNγ(γ-干扰素)对TRAIL(肿瘤坏死因子相关凋亡诱导配体)诱导神经母细胞瘤细胞株SMS-KCNR(KCNR)细胞凋亡的影响及其发生机制。方法应用RT-PCR方法检测IFNγ作用前后KCNR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+Caspase8抑制剂(zIETD-FMK)+TRAIL对KCNR细胞生长及凋亡的影响;应用比色法测定Caspase8相对活性。结果KCNR细胞不表达Caspase8,IFNγ作用48h后的KCNR细胞Caspase8表达明显增加;KCNR细胞对TRAIL不敏感,IFNγ诱导表达Caspase8的KCNR细胞对TRAIL敏感;IFNγ+TRAIL组Caspase8相对活性明显高于TRAIL组及抑制剂组;zI-ETD-FMK能阻断Caspase8的活化而抑制TRAIL对KCNR细胞的诱导凋亡作用。结论IFNγ通过诱导Caspase8表达而逆转KCNR细胞对TRAIL诱导凋亡的耐受。
Objective To investigate the effect and mechanism of IFNγ (γ-interferon) on the apoptosis of neuroblastoma cell line SMS-KCNR (KCNR) induced by TRAIL (tumor necrosis factor-related apoptosis inducing ligand). Methods The expression of Caspase8 in KCNR cells was detected by RT-PCR. IFNγ, TRAIL, IFNγ + TRAIL and IFNγ + Caspase8 were detected by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) Inhibitors (zIETD-FMK) + TRAIL on the growth and apoptosis of KCNR cells; the relative activity of Caspase8 was determined by colorimetric assay. Results KCNR cells did not express Caspase8, but Caspase8 expression of KCNR cells increased significantly after IFNγ treatment for 48 hours. KCNR cells were not sensitive to TRAIL. IFNγ induced Caspase8-expressing KCNR cells to TRAIL. Relative activity of Caspase8 in IFNγ + TRAIL group was significantly higher than that in TRAIL group Inhibitor group; zI-ETD-FMK can block the activation of Caspase8 and inhibit TRAIL on KCNR cells induced apoptosis. CONCLUSION: IFNγ can reverse the tolerance of KCNR cells to TRAIL-induced apoptosis by inducing the expression of Caspase8.