多柔比星温热化疗诱导A549细胞凋亡实验研究  

Study of Doxorubicin on Induction A549 Cells Apoptosis by Hyperthermic Chemotherapy

作　　者：张继贤[1] 赵莺[1] 张启昆[2] 魏海东[2] 

机构地区：[1]浙江省余姚市人民医院呼吸科,315400 [2]兰州大学第二医院呼吸科,730030

出　　处：《医药导报》2007年第8期868-871,共4页Herald of Medicine

摘　　要：目的研究多柔比星(DRB)温热化疗诱导人肺腺癌A549细胞凋亡的效果,探讨其作用机制。方法对照组A549细胞不经处理,37℃培养箱中常规培养;化疗组A549细胞分为4组,DRB浓度分别为0.5,1.0,2.0和4.0μg.mL-1,均37℃培养箱中常规培养;热疗组A549细胞采用水浴加温法,温度设定为42.5℃,加热时间30 min,然后在37℃孵箱中培养;DRB温热化疗组(热疗+化疗)DRB浓度同化疗组,处理方法同热疗组。四氮甲唑蓝(MTT)比色法测定各组细胞抑制率,流式细胞仪(FCM法)测定细胞内DRB荧光强度及Fas蛋白水平变化,比色法检测细胞内Caspase-3活性变化。结果DRB温热化疗可明显增强DRB对A549细胞的抑制作用并诱导细胞凋亡;FCM分析显示,化疗组、DRB温热化疗组细胞内DRB浓度升高,且呈浓度依赖性;DRB温热化疗组细胞内DRB浓度高于化疗组(P<0.01)。细胞周期分析显示,化疗组和DRB温热化疗组G1期细胞所占比例降低,S期细胞比例增高,细胞被阻滞于S期。化疗组和DRB温热化疗组Fas蛋白表达上调,Caspase-3活性增强。结论DRB温热化疗可增加A549细胞凋亡,其机制可能是通过激活Fas依赖的Caspase-3诱导凋亡起作用。Objective To investigate the effects of hyperthermic chemotherapy of doxorubicin （DRB） on inducing apoptosis of lung adenocarcinoma A549 cells and explore its possible mechanisms. Methods A549 cells without treatment were cultured at 37℃ in a normal way as the control group. A549 cells in the chemotherapeutic groups were treated with DRB at doses of 0.5,1.0,2.0 and 4.0 μg ·mL^-1, respectively, and cultured in a normal way as well. A549 cells in the hyperthermic group were heated in a water bath at 42.5℃ for 30 min, and then cultured at 37 ℃ in a normal way. A549 cells in the hyperthermic chemotheraputy groups were treated with DRB at the same dose with those in the chemotherapeutic group and hyperthermic treated in the same way as those in the hyperthermic group. MTT assay was used to detect cells suppression rate in each group, flow cytometry was employed to detect the fluorescence intensity of DRB in the cells and expression levels of Fas antigens, colorimetric assay was employed to detect the activation of Caspase-3. Results The hyperthermic chemotherapy significantly increased the inhibition of A549 cells by DRB and indued cell apoptosis. FCM shown that the concentration of DRB in both chemotherapy groups and hyperthermic chemotherapy groups was increased in a concentration-dependent way, and the concentration of DRB in hyperthermic chemotherapy groups was significantly higher than that in chemotherapy groups （ P 〈 0.01 ）. Cell cycle analysis indicated that decreased G1 phase and increased S phase proportion, as well as apparent S phase arrest were shown in chemotherapy groups, and DRB. The expression of Fas was increased significantly and Caspase-3 was also activated in both chemotherapy groups and hyperthermic chemotherapy groups. Conclusion Hyperthermic chemotherapy of DRB could induce A549 cell apoptosis significantly, the mechanism of which may be through the pathway by activating Caspase- 3 in a Fas-depended way.

关 键 词：多柔比星 温热化疗 凋亡 

分 类 号：R979.1[医药卫生—药品]