小剂量甲氨蝶呤联合GM-CSF对U937细胞的诱导分化作用  

Low dose methotrexate combined with GM-CSF induced differentiation of U937 cell s

作　　者：陈静[1] 顾龙君[1] 应大明[1] 

机构地区：[1]上海第二医科大学附属新华医院,上海儿童医学中心,200127

出　　处：《中华血液学杂志》2003年第8期430-432,共3页Chinese Journal of Hematology

基　　金：卫生部科研基金 ( 98 2 30 3);上海市科学技术发展基金资助项目 ( 984 1190 10 )

摘　　要：目的　探索甲氨蝶呤 (MTX)对急性单核细胞白血病的药效。方法　以U937细胞株为实验模型 ,以细胞形态学、NBT还原实验及流式细胞术检测CD1 4 阳性细胞为评价U937细胞分化的指标 ,以TRAP ELISA试剂盒检测端粒酶活性。结果　经 2 0nmol LMTX作用 2 4 ,48,72h后 ,U937细胞体积明显增大 ,核 浆比例逐渐缩小 ;NBT还原实验逐渐呈现阳性 ;培养 72h后 ,CD1 4 阳性细胞从 3 %上升到2 0 % ,提示出现部分分化。单用 1 0 0U mlGM CSF并未对U937细胞产生诱导分化作用 ,但当此剂量的GM CSF与 2 0nmol LMTX同时作用于U937细胞 72h后 ,63 %的细胞出现分化。MTX联合GM CSF作用2 4 ,48,72h后随着细胞被诱导分化 ,端粒酶活性逐渐从 2 .1 1 (未用药前 )分别下降到 1 .48,0 .77和 0 .2 4。结论　小剂量MTX联合GMObjective To explore the effect of methotrexate on acute monoblastic leukemia cells, and the potential role of low-dose methotrexate in the treatment of monoblastic le ukemia. Methods By using U937 cell line as a model, the induced differentiation of MTX or GM-CSF or their combination was detected by NBT reduction test, morphological observation and CD 14 expression. TRAP-ELISA was used for measuring telomerase activity. Results After incubation with 20 nmol/L methotrexate for 24,48,72 and 96 hours, the si ze of the U937 cells increased, the nuclear/plasma ratio gradually decreased, NB T reductive test became positive, and CD 14 positive cells increased from 3 % to 20% after 72 hours incubation, indicating partial differentiation. Treatmen t of U937 cells with 100?U/ml GM-CSF alone failed to induce differentiation. H owever, GM-CSF combined with low-dose methotrexate resulted in 63% of differen tiated cells after 72 hours incubation, and telomerase activity of the U937 cell s decreased from 2.11(before treatment) to 1.48, 0.77, 0.24 (after treatment f or 24,48,72 hrs respectively). The extent of the decrease of telomerase activity was in proportion with the degree of differentiation. Conclusion Monoblastic leukemia might be treated with low-dose methotrexate plus GM-CSF as a differentiation induction regimen.

关 键 词：急性单核细胞白血病 U937细胞 甲氨蝶呤 粒细胞-巨噬细胞集落刺激因子 细胞分化 

分 类 号：R733.71[医药卫生—肿瘤]