【摘要】 目的探讨四嗪二甲酰胺(ZGDHu-1)对慢性粒细胞白血病细胞株K562(K562-S)和耐药细胞株K562/Adr(K562-R)的抑制作用及其对p210 BCR/Ab l融合蛋白和转录核因子κB(NF-κB)表达的影响。方法 MTT比色法观察其对K562-S细胞增殖的抑制作用,采用Hoechst 33258荧光染色检测细胞凋亡,流式细胞仪及细胞克隆形成初步分析其对白血病干细胞的影响,通过Western印迹分析PARP-1、p210 BCR/Abl、c-Abl、NF-κB的表达。结果不同浓度的ZGDHu-1对K562-S细胞均有增殖抑制作用,呈现剂量-时间依赖关系,ZGDHu-1作用48h时的IC50值为0.25μg/ml。在ZGDHu-1作用7天后,K562-S及K562-R细胞克隆形成被完全抑制。ZGDHu-1作用48h后,出现PARP-1裂解,诱发细胞凋亡,下调K562-R细胞p210 BCR/Abl及c-Abl的表达,抑制K562-S及K562-R细胞质内NF-κB/p65的表达。结论 ZGDHu-1能够抑制K562-S及耐药细胞株K562/Adr(K562-R)细胞的增殖,促进细胞凋亡。通过抑制p210 BCR/Abl的表达,降低NF-κB的活性,阻断NF-κB信号传导通路,尤其对于耐药型白血病细胞具有广泛的应用价值。更多还原

【Abstract】 Objective To explore the effect of ZGDHu-1 on proliferation of chronic myeloid leukemia cell K562(K562-S)and its multidrug resistant cell line K562/Adr(K562-R),and to discuss the expression of the p210 BCR/Abl and NF-κB.Methods MTT was used to determine the proliferative effects of ZGDHu-1 on K562-S cell.K562-S and K562-R cell suspension was stained with Hoechst33342.The leukemia stem cells was preliminary analyzed by flow cytometry and colon forming.Western blotting was used to examine the expression of the PARP-1,p210 BCR/Abl and NF-κB.Results ZGDHu-1 could inhibit K562-S cell proliferation within different concentration,which be presented in dose and time dependent manner.Treated with ZGDHu-1 for 48 hours,the K562-S cell IC50 value was 0.25μg/ml.The K562-S and K562-R cells apoptosis was confirmed by Hoechst33258 straining.Treated with ZGDHu-1 after 7 days,K562-S and K562-R cell colons forming was absolutely inhibited.After incubating with ZGDHu-1 for 48 hours,the cleavage of PARP was observed in the process of apoptosis of K562-S cell and K562-R cell,and the protein expressions of p210BCR/Abl,c-Abl and NF-κB/p65 were all decreased.Conclusion ZGDHu-1 can inhibit K562-S and K562-R cell proliferation and induce apoptosis.The mechanism may be associated with inhibiting the expression of p210 BCR/Abl and the blockage of NF-κB signal transduction pathway.ZGDHu-1 may have widespread application value in treatment of drug resistance to chemotherapy in the patients with leukemia.更多还原