人膀胱癌细胞系SW780中侧群细胞的功能鉴定  

Identification of side population cells from bladder cancer cells by DyeCycle Violet staining

作　　者：佘军军[1] 石钊琪[1] 薛孝斌[1] 王红军[1] 车向明[1] 

机构地区：[1]西安交通大学医学院第一附属医院普通外科,西安710061

出　　处：《山西医科大学学报》2013年第10期776-780,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81072108)

摘　　要：目的使用DCV荧光染料从SW780人膀胱癌细胞系中分离侧群细胞(SP细胞),并观察该侧群细胞是否具有肿瘤干细胞样生物学特征。方法 SW780细胞体外培养,用DCV荧光染料分析SP细胞比率。分选SP与NSP细胞群落后体外培养扩增。软琼脂克隆集落生成实验比较SP细胞与NSP细胞的体外单克隆生长能力。采用RT-PCR法比较ABCG2、MDR1、Oct-4及Bmi-1等基因表达差异。结果 SW780细胞的SP比率为1.6%。经体外培养后SP细胞可以产生SP细胞与NSP细胞,NSP细胞只能产生NSP细胞。SP细胞体外单克隆生长率显著大于NSP细胞(P<0.01)。半定量RT-PCR结果表明SP细胞的ABCG2、MDR1、Oct-4及Bmi-1基因表达显著高于NSP细胞。结论流式细胞仪激发DCV荧光染料可从人膀胱癌SW780细胞系中分离出侧群细胞,该侧群细胞具有肿瘤干细胞样生物学特征。Objective To isolate SP cells from SW780 bladder cancer cell lines using DCV staining and to explore whether SP cells have stem cell-like characteristics.Methods SW780 cell lines were cultured in vitro.SP cells were sorted from SW780 cell lines and the percentage was analyzed using DCV staining.Both SP cells and NSP cells were cultured in vitro after sorting.The colony-forming ability of SP cells and NSP cells were compared using soft agar assay.The expression of ABCG2,MDR1,Oct-4 and Bmi-1 was detected and compared between SP and NSP cells using RT-PCR.Results The percentage of DCV SP cells was 1.6％ in SW780 cell lines.SP cells could generate SP cells and NSP cells,while NSP could only generate NSP cells.The colony-forming ability of SP cells was significantly higher than NSP cells （P ＜ 0.01）.RT-PCR results showed that the expression levels of ABCG2,MDR1,Oct-4 and Bmi-1 were higher in SP cells than those in NSP cells.Conclusion The results suggest that DCV staining could be used to isolate and identify SP cells from cancer cell lines,and these SP cells have stem cell-like biological characteristics.

关 键 词：膀胱癌 SW780细胞系 肿瘤干细胞 侧群细胞 

分 类 号：R737.14[医药卫生—肿瘤]