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目的 探讨重组人肿瘤坏死因子 α(rhTNF α)对白血病细胞的作用及其机制 ,寻求白血病免疫疗法的理论依据。方法 以早幼粒白血病细胞株HL 60细胞为研究对象 ,利用苏木素染色及免疫细胞化学的方法 ,观察rhTNF α对HL 60细胞凋亡及其bcl 2基因表达的影响。结果 10U/ml、10 0U/ml和 5 0 0U/mlrhT NF α与HL 60细胞共同培养 2～ 2 4h ,除 10U/ml培养 2h组外余各组凋亡细胞数均明显高于空白对照组 (P<0 .0 5 ) ,bcl 2基因阳性表达细胞数均明显低于空白对照组 (P <0 .0 5 ) ,二者呈明显负相关。随rhTNF α浓度增高 ,其作用增强 ,8h为作用的高峰时间。结论 rhTNF α能促进HL 60细胞的凋亡 ;抑制凋亡调控基因bcl 2的表达可能是TNF α促进HL 60细胞凋亡的机制之一。
Objective To investigate the effect and mechanism of recombinant human tumor necrosis factor α (rhTNFα) on leukemia cells, and to seek theoretical basis for leukemia immunotherapy. Methods HL 60 cells of promyelocytic leukemia cell line were studied. The effects of rhTNF α on the apoptosis of HL 60 cells and the expression of bcl 2 gene were observed by hematoxylin staining and immunocytochemistry. RESULTS: 10 U/ml, 100 U/ml and 500 U/ml rhT NF alpha were co-cultured with HL 60 cells for 2 to 24 hours. The number of apoptotic cells in each group was significantly higher than that of the control group except for 10 U/ml cultured for 2 hours. (P<0.05), the number of bcl-2 positive cells was significantly lower than that of the blank control group (P<0.05). There was a significant negative correlation between the two. With the increase of rhTNFα concentration, its effect was enhanced and 8h was the peak time of action. Conclusion rhTNFα can promote the apoptosis of HL 60 cells; inhibiting the expression of bcl 2 may be one of the mechanisms of TNF α promoting apoptosis of HL 60 cells.