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目的 :观察 Fas对 HL - 6 0细胞的异常增殖是否有调节作用。方法 :用基因重组技术将 Fas的细胞外区跨膜区与 IL -6信号传导子 gp130细胞内区构成嵌合型受体 (Fas/130 ) ,同时 ,将 Fas死亡区域 (Fas DD)替换 Fas/130中 130细胞内区无结构域部分 (Fas/130 f) ,分别在 HL - 6 0中表达后 ,用抗 Fas抗体激活这些嵌合型受体 ,随后用免疫组化和蛋白质印迹法分析受体细胞内区形成同源性三聚体 (130 cyt- 130 cyt- 130 cyt及 Fas DD- Fas DD- Fas DD)后细胞内 Stat3的表达及磷酸化。结果 :转染p ED Fas/130后用抗 Fas抗体作用 6～ 8h,HL - 6 0细胞 Stat3表达下降 (P<0 .0 5 ) ,而 Fas/130 f组的 Stat3的表达下降更明显(P<0 .0 1) ;两实验组中 Fas/130 f组在特异性抗体诱导 10 m in时 ,Stat3 Tyr70 5磷酸化较 Fas/130组明显降低 (P<0 .0 1)。结论 :Fas死亡域抑制白血病 HL- 6 0细胞内 Stat3的表达 ,并同时抑制 Stat3Tyr70 5磷酸化。
Objective: To observe whether Fas can regulate the abnormal proliferation of HL - 60 cells. Methods: Fas / 130 was constructed by transmembrane domain of Fas in extracellular domain and intracellular domain of IL-6 signaling gp130 by gene recombination technique. At the same time, the Fas death domain (Fas DD) was replaced by Fas / 130 in the intracellular domain without domain portion (Fas / 130 f), respectively, after expression in HL - 60, the chimeric receptors were activated with anti - Fas antibodies, followed by immunohistochemistry and Western blot analysis The expression and phosphorylation of Stat3 in the cells after homologous trimer (130 cyt-130 cyt-130 cyt and Fas DD-Fas DD-Fas DD) formed in the receptor intracellular domain. Results: The expression of Stat3 in HL - 60 cells decreased (P <0.05) after anti - Fas antibody treatment for 6 h and 8 h after transfection with p ED Fas / P <0.01). Stat3 Tyr70 5 phosphorylation of Fas / 130 f group was significantly lower than that of Fas / 130 group (P <0.01) at 10 mins induction of specific antibody in both experimental groups. Conclusion: The Fas death domain inhibits the expression of Stat3 in leukemia HL-60 cells and inhibits the phosphorylation of Stat3Tyr70 5.