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目的研究二氟甲基鸟氨酸(difluromethylornithine,DFMO)对人T淋巴瘤Jurkat细胞的影响及其作用机制,为探讨DFMO能否用于治疗人白血病提供实验依据。方法DFMO(0～10mmol/L)处理人T淋巴瘤Jurkat细胞24～72h后,MTS法检测细胞的存活率,化学分析法检测精胺氧化酶(spermine oxidase,SMO)和乙酰多胺氧化酶(acetylpolyamine oxidase,APAO)活性,DNA片段化分析检测细胞凋亡,荧光染料法检测细胞线粒体膜电位变化,Western blot法检测细胞内Bax含量,分光光度法检测caspase-3酶活性。结果DFMO处理细胞能显著抑制Jurkat细胞的生长(P<0.01),抑制率随药物浓度的增加和作用时间的延长而加大,DFMO浓度为10mmol/L时,24h抑制率为33%,48h抑制率为38%,72h抑制率为49%。DFMO处理可导致Jurkat细胞DNA片段化,促进Bax由细胞质向线粒体转移,细胞内caspase-3活性增加(增加46%,P<0.05),并伴有线粒体膜电位的显著降低,DFMO浓液为2和5mmol/L时,分别降低29%和53%(P<0.01)。DFMO还能诱导细胞内SMO活性(增加1.1倍,P<0.05)但对APAO的活性无诱导性影响。结论DFMO对人T淋巴瘤Jurkat细胞具有生长抑制和诱导凋亡的作用。
Objective To investigate the effect and mechanism of difluromethylornithine (DFMO) on human T lymphoma Jurkat cells and to provide an experimental basis for investigating whether DFMO could be used to treat human leukemia. Methods The survival rate of human T lymphoma Jurkat cells treated with DFMO (0 ~ 10mmol / L) for 24 ~ 72h was determined by MTS method. The levels of spermine oxidase (SMO) and acetyl polyamine oxidase acetylpolyamine oxidase (APAO) activity, DNA fragmentation analysis was used to detect apoptosis. Fluorescent dye method was used to detect the change of mitochondrial membrane potential. Western blot was used to detect the content of Bax. Spectrophotometry was used to detect the activity of caspase-3. Results DFMO treatment significantly inhibited the growth of Jurkat cells (P <0.01). The inhibition rate increased with the increase of drug concentration and prolongation of action time. DFMO inhibited the growth of Jurkat cells at the concentration of 10 mmol / L for 33 h and at 48 h The rate was 38%, 72h inhibition rate of 49%. DFMO treatment led to DNA fragmentation in Jurkat cells, promoted the transfer of Bax from cytoplasm to mitochondria, and an increase in intracellular caspase-3 activity (46% increase, P <0.05) accompanied by a significant decrease in mitochondrial membrane potential with DFMO concentrate at 2 And 5 mmol / L decreased by 29% and 53%, respectively (P <0.01). DFMO also induced intracellular SMO activity (1.1-fold increase, P <0.05) but had no inducing effect on APAO activity. Conclusion DFMO can inhibit the growth of human T lymphoma Jurkat cells and induce apoptosis.