硫酸软骨素对HL60细胞增殖分化的影响(英文)  

Effect of chondroitin sulfate on the proliferation and differentiation of HL60 cells

作　　者：徐文华[1] 虞浩[1] 张银亭[1] 葛银林[1] 耿芳宋[1] 

机构地区：[1]青岛大学医学院生物化学与分子生物学教研室,山东省青岛市266021

出　　处：《中国临床康复》2006年第37期166-168,F0003,共4页Chinese Journal of Clinical Rehabilitation

基　　金：青岛市科技局科技计划项目(03-1-YN-14)~~

摘　　要：背景:硫酸软骨素是细胞基质的重要成分,可促进肿瘤细胞的增殖,抑制其转移。目的:观察硫酸软骨素对阿霉素作用下HL60细胞增殖分化的影响。设计:开放性实验。单位:青岛大学医学院生物化学与分子生物学教研室。材料:实验于2003-09/2004-12在青岛大学医学院生物化学与分子生物学研究室完成。HL60细胞株购于中国医学科学院上海细胞库,为人类早幼粒白血病细胞。牛软骨硫酸软骨素(Sigma)。方法:①细胞传代培养后将进入对数增殖期的细胞用含体积分数0.1灭活胎牛血清的RPMI1640培养液配制成1×108L-1的细胞悬液,分装于45个培养瓶中,4mL/瓶。②取分装好的细胞悬液15瓶,按照0,5,25,50,75mg/L浓度加入硫酸软骨素,每个浓度平行3瓶,空白对照组加入等量的0.01mol/L磷酸盐缓冲液。采取细胞计数法测定硫酸软骨素处理后的HL60细胞密度的变化。③取分装好的细胞悬液30瓶,分为硫酸软骨素+阿霉素组、硫酸软骨素组,各15瓶。按照0,5,25,50,75mg/L浓度向两组加入硫酸软骨素,每个浓度平行3瓶,空白对照组加入等量的0.01mmol/L磷酸盐缓冲液。用MTT法测定硫酸软骨素处理后的HL60细胞加入阿霉素后细胞存活率的变化。④取分装好的细胞悬液15瓶,按照0,5,25,50,75mg/L浓度加入硫酸软骨素,每个浓度平行3瓶,空白对照组加入等量的0.01mol/L磷酸盐缓冲液。用酶联免疫反应测定各组细胞的酸性磷酸酶活性,观察其对细胞分化的影响。主要观察指标:①硫酸软骨素对HL60细胞增殖的影响。②硫酸软骨素+阿霉素对HL60细胞存活率的影响。③硫酸软骨素对HL60细胞酸性磷酸酶活性的影响。结果:共制备细胞悬液45瓶,全部进入结果分析。①与空白对照组比较,不同浓度硫酸软骨素处理24h后HL60细胞密度均显著升高(P<0.01);且50,75mg/L硫酸软骨素浓度组的细胞密度升高尤为明显,但两组间比较基本相似(P>0.05),说明在�BACKGROUND： Chondroitin sulfate is the important component of cell matrix, it can accelerate the proliferation of tumor cells and restrain its transfer. OBJECTIVE： To observe the effects of chondroitin sulfate on the proliferation and differentiation of HL60 cells under the action of adriamycin. DESIGN： An open experiment. SETTING： Department of Biochemistry and Molecular Biology, Medical College of Qingdao University. MATERIALS： The experiments were carried out in the Research Room of Biochemistry and Molecular Biology, Medical College of Qingdao University of September 2003 to December 2004. Experimental materials and reagents： HL60 cell strains, which were the cells from promylocytic leukemia, were purchased from Shanghai Cell Bank, Chinese Academy of Medical Sciences; Bovine cartilage chondroitin sulfate （Sigma） was also USed. METHODS： ①After the passage and culture, the cells at the logarithmic proliferative phase were dispensed into cell suspension of 1×10^8 L^-1 with RPMI1640 culture medium containing inactivated fetal bovine serum of 0.1 in volume fraction, and then filled into the culture bottles with 4 mL in each bottle for a total of 45 bottles. ② Chondroitin sulfate was added to 15 bottles filled with cell suspension according to the concentrations of 0, 5, 25, 50 and 75 mg/L respectively, and 3 bottles for each concentration, and 0.01 mol/L phosphate buffer （pH7.2） was added in the blank control group. Then the density of HL60 cells was determined by cell counting after treatment of ehondroitin sulfate. ③ Thirty bottles filled with cell suspension were divided into ehondroitin sulfate±adriamycin group and chon- droitin sulfate group, 15 bottles in each group. Chondroitin sulfate of 0, 5, 25, 50 and 75 mg,/L was added to the two groups, and 3 bottles for each concentration, and 0.01 mol/L phosphate buffer （pH7.2） wass adokd in the blank control group. Then the survival rate of ehondroitin sulfate treated HL60 was detected after adding adriamycin. ④Chondroit

关 键 词：细胞增殖 细胞计数 HL-60细胞 软骨素硫酸盐类 多柔比星 

分 类 号：R349.2[医药卫生—基础医学]