吉西他滨对CD34^+ CD38^-髓系白血病干细胞的增殖抑制和凋亡诱导作用  

Effects of gemcitabine on inhibiting proliferation and inducing apoptosis of CD34^+ CD38^- myeloid leukemia stem cells

作　　者：韩慧娟[1] 胡亮杉[2] 邓兰[1] 高莹莹[1] 郭坤元[1] 

机构地区：[1]南方医科大学珠江医院血液内科,广东广州510282 [2]广东省第二人民医院检验科,广东广州510317

出　　处：《中国病理生理杂志》2013年第3期541-545,548,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30973454)

摘　　要：目的:探讨与阿糖胞苷(Ara-C)结构相似的新型脱氧胞苷相似物和核苷还原酶抑制剂——吉西他滨(gemeitabine,GEM)对CD34+CD38-KG1a髓系白血病干细胞(LSCs)增殖抑制和诱导凋亡的影响。方法:流式细胞术检测急性髓系白血病KG1a细胞表面CD34和CD38的表达;24 h和持续用药软琼脂克隆形成实验观察不同浓度GEM对KG1a细胞增殖的影响,流式细胞术检测不同浓度GEM对KG1a细胞周期的影响,Annexin V/PI双染法检测不同浓度GEM对KG1a细胞凋亡的影响。结果:急性髓系白血病KG1a细胞中CD34+CD38-占(98.02±0.72)%。0.05 mg/L、0.1 mg/L和0.5 mg/L的GEM分别作用KG1a细胞24 h、48 h和72 h后,0.5 mg/L GEM作用KG1a细胞24 h后,G0/G1期细胞高于盐水对照组(P<0.05),而0.05 mg/L和0.1 mg/L GEM作用KG1a细胞24 h后,G0/G1期细胞与盐水对照组相比,差异无统计学意义(P>0.05)。0.05 mg/L、0.1 mg/L和0.5 mg/LGEM作用KG1a细胞24 h后,软琼脂培养第14 d和21d后,0.1 mg/L和0.5 mg/L组形成的克隆数,低于盐水对照组(P<0.05),0.05 mg/L GEM组14 d、21 d的克隆数与对照组相比,差异无统计学意义(P>0.05)。0.05 mg/L、0.1 mg/L、0.5 mg/L GEM和Ara-C持续作用组,软琼脂培养14 d和21d均未见集落生长,与对照组相比差异显著(P<0.05)。0.05 mg/L、0.1 mg/L GEM作用KG1a细胞后其凋亡率与盐水对照组相比,差异无统计学意义(P>0.05),而0.5 mg/L GEM作用24 h后KG1a细胞凋亡率显著高于盐水对照组(P<0.05)。结论:GEM能抑制CD34+CD38-髓系白血病干细胞增殖和克隆形成,并将CD34+CD38-KG1a细胞阻滞在G0/G1期和诱导其凋亡。AIM ： To investigate the effects of gemcitabine （ GEM）, a novel analog of deoxycytidine and nucle- oside reductase inhibitor similar to cytarabine （Ara-C） in structure, on the proliferation and apoptosis of myeloid leukemic stem cells （LSCs）, CD34＋CD38- KGla cells. METHODS： The expression of CD34 and CD38 on the surface of acute myeloid leukemia KG1 a cells was detected by flow cytometry. The effects of GEM at various concentrations for 24 h and sustained medication for 14 d and 21 d on the proliferation and colony-forming ability of KGla cells were analyzed by soft agar colony-forming experiment. The changes of the cell cycle of KGla cells treated with various concentrations of GEM were tested by flow cytometry. The apoptosis of KG1 a cells was determined by flow cytometry with the staining of Annexin V-FITC and propidium iodide （PI）. RESULTS： The percentage of CD34 ~ CD38- cells in acute myeloid leukemia KGla cells was （98.02 ± 0.72） %. Treatments with 0. 05 mg/L and 0.1 mg/L GEM for 24 h were similar to saline control group in cell cycle distribution of the KGla cells, whereas KGla cells treated with 0.5 mg/L GEM for 24 h were arrested at G0/G1 phase. After treatment with 0.1 mg/L and 0.5 mg/L GEM for 24 h, the colony numbers at 14 d and 21 d were low- er than that in saline control group. No difference of the colony numbers between the cells treated with normal saline and 0. 05 mg/L GEM for 14 d and 21 d was observed. After sustained medication with 0. 05 mg/L, 0.1 mg/L and 0.5 mg/L GEM and Ara-C for 14 d and 21 d, the colony numbers decreased as compared to saline control group. Treatment with 0. 5 mg/L GEM for 24 h increased the apoptotic rate of KG1 a cells compared with saline control group, while treatments with O. 05 mg/L and 0.1 mg/L GEM for 24 h were similar to saline control group in cell apoptosis. CONCLUSION ： GEM in- hibits the proliferation and colony-forming ability, arrests the cell cycle at G0/G1 phase and induces apoptosis of CD34＋CD38- acute myeloid leukemia

关 键 词：吉西他滨 KG1a细胞 细胞周期 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤]