表柔比星对人骨肉瘤干细胞分离纯化的影响  

Influence of epirubicin on human osteosarcoma stem cell isolation and purification

作　　者：杨寒石 吕游[1] 陈景阳[1] 袁峰[2] 郭开今[2] 葛保健[2] 

机构地区：[1]徐州医学院研究生院,221004 [2]徐州医学院附属医院骨科

出　　处：《中华实验外科杂志》2015年第7期1560-1562,共3页Chinese Journal of Experimental Surgery

基　　金：徐州市科学技术局资助项目（xzzd1146）

摘　　要：目的 探讨分离纯化人骨肉瘤干细胞的新途径.方法 体外培养人骨肉瘤U2OS细胞和人骨肉瘤干细胞,免疫荧光检测人骨肉瘤干细胞球中CD105表达.表柔比星处理人骨肉瘤干细胞球,细胞计数试剂盒（CCK-8）法测增殖活性.Western blot法检测表柔比星处理后人骨肉瘤干细胞球Nanog和Oct4蛋白表达.结果 荧光显微镜下可见,人骨肉瘤干细胞球CD105呈阳性表达.表柔比星处理后人骨肉瘤干细胞球生长受抑制.Western blot检测结果显示,2.5 mg/L表柔比星处理组Nanog蛋白和Oct4蛋白相对表达量均为最高且不再增加.2.5 mg/L表柔比星为分离培养人骨肉瘤干细胞的最适浓度.结论 悬浮培养联合表柔比星较传统无血清培养,能更好的分离、纯化人骨肉瘤干细胞.Objective To explore the new approach in separation and purification of human osteosarcoma stem cells.Methods Human osteosarcoma U2OS cells and osteosarcoma stem cells were cultured in vitro.The expression of CD105 in osteosarcoma stem cell spheres was detected using immunofluorescence assay.The osteosarcoma stem cell spheres were treated with epirubicin,and their proliferative activities were determined using cell counting kit-8 （CCK-8）.The expression of Nanog and Oct4 proteins in osteosarcoma stem cells treated with epirubicin was detected using Western blotting.Results Fluorescence microscope showed that the expression of stem cell CD105 in osteosarcoma stem cell spheres was positive.The results of CCK-8 showed that growth inhibition for osteosarcoma stem cells treated with epirubicin was increased.According to the results of Western blotting,the relative expression level of Nanog protein and Oct4 protein for osteosarcoma stem cells in the group treated with 2.5 mg/L epirubicin reached the peak.2.5 mg/L of epirubicin was the optimal concentration for isolation and culture of human osteosarcoma stem cells.Conclusion Suspension culture combined with epirubicin can better isolate and purify human osteosarcoma stem cells than the traditional serum-free culture.

关 键 词：骨肉瘤 骨肉瘤干细胞 表柔比星 培养 

分 类 号：R738.1[医药卫生—肿瘤]