热疗联合柔红霉素对CD34^＋CD38^-白血病细胞增殖和凋亡的影响  

Effects of hyperthermia plus daunorubicin on the proliferation and apoptosis of CD34^+CD38^-leukemia cells

作　　者：王国征[1] 郭坤元[1] 孙明[1] 佘妙容[2] 胡亮杉[1] 李章球[1] 

机构地区：[1]南方医科大学珠江医院血液科,广州510282 [2]广东省人民医院血液科

出　　处：《解放军医学杂志》2009年第4期419-422,共4页Medical Journal of Chinese People's Liberation Army

基　　金：广东省自然科学基金资助项目(7001196)

摘　　要：目的观察热疗联合柔红霉素(DNR)对CD34+CD38-急性髓白血病KG1a细胞增殖的抑制作用和对其凋亡的影响。方法免疫磁珠分离KG1a细胞中的CD34+CD38-细胞,以DNR作用48h后抑制50%CD34+CD38-细胞生长的药物浓度(IC50)作为后续实验的工作浓度。将CD34+CD38-细胞分为对照组、热疗组(40℃、42℃)、化疗组和热化疗组(40℃、42℃)。热疗、热化疗组在恒温水浴箱(40℃或42℃)中加热60min,化疗组、热化疗组以IC50浓度的DNR处理48h。MTT法检测DNR对CD34+CD38-KG1a细胞增殖的抑制作用;流式细胞仪检测细胞凋亡率及42℃条件下细胞内DNR平均荧光强度的变化;甲基纤维素培养体系分析42℃条件下细胞的克隆形成能力。结果KG1a细胞中分离出的CD34+CD38-细胞纯度达95%以上,DNR的IC50为0.40μg/ml。热疗组、化疗组、热化疗组CD34+CD38-细胞增殖均明显受抑,且抑制作用随温度升高而增强(P<0.05)。热疗组、化疗组及热化疗组细胞凋亡率均较对照组显著增加,且随温度升高凋亡率增加(P<0.05),热化疗组较化疗组及相同温度热疗组的细胞凋亡率显著增加(P<0.05)。42℃热化疗组细胞内的DNR荧光强度(6.74±0.58)明显高于对照组(0.26±0.05)及化疗组(2.08±0.14,P<0.05)。42℃热疗组、化疗组、42℃热化疗组细胞的集落形成率明显低于对照组,且42℃热化疗组明显低于42℃热疗组和化疗组(P<0.05)。结论热疗能增强DNR抑制CD34+CD38-KG1a细胞增殖的作用,使细胞凋亡增多。Objective To explore the effects of hyperthermia plus daunorubicin （DNR） on the proliferation and apoptosis of acute myelogenous leukemia CD34^＋ CD38^- cells, Methods CD34^＋ CD38^- cells were isolated from KGla cells with magnetic activated cell sorting （MACS）. 50% inhibiting concentration （IC50） of DNR against CD34^＋ CD38^- cell was assessed by MTT assay 48 hours after treating with DNR, and it was taken as the concentration for the subsequent experiments. CD34^＋ CD38^- cells were divided into control group, 40℃ or 42℃ hyperthermia groups, chemotherapy group, and 40℃ or 42℃ hyperthermia plus chemotherapy group. CD34^＋ CD38^- cells in hyperthermia group and hyperthermia plus chemotherapy groups were heated at 40℃ or 42℃ in a water bath for 60 minutes. The inhibition of proliferation of CD34^＋ CD38^- cells by DNR was evaluated by MTT assay, and apoptosis rates as well as fluoscence intensity of DNR inside CD34^＋ CD38^- cells were assessed by flow cytometry. Moreover, clonality was analyzed in methylcellulose system at 42℃. Resuits The purity of CD34^＋ CD38^- cells isolated from KGla was 95% and higher, and the IC50 of CD34^＋ CD38^- cells against DNR was 0. 40μg/ml. As compared with the control group, the proliferation of CD34^＋ CD38^- cells was inhibited in all the other groups, and the inhibitive effects were positively dependent on temperature （P〈0. 05）. As compared with the control group, the apoptosis rates were higher in all the other groups, and the elevation of apoptosis rate was positively depent on temperature （P〈0. 05）. Moreover, the apoptosis rate was higher in hyperthermia plus chemotherapy group than in both chemotherapy group and hyperthermia group with the same temperature （P〈0. 05）. At 42℃, the fluorescence intensity of DNR in hyperthermia plus chemotherapy group （6. 74±0. 58） was significantly higher than that in both control group （0. 26±0. 05） and chemotherapy group （2. 08±0. 14, P〈0. 05）. Compared to the control

关 键 词：白血病 髓样 柔红霉素 高温 诱发 肿瘤干细胞 细胞凋亡 

分 类 号：R733.712[医药卫生—肿瘤]