摘要目的 检测人神经母细胞瘤细胞系SH-SY5Y及WN中是否存在侧群(side population,SP)细胞,并研究该SP细胞是否具有干细胞某些生物学特性.方法 经Hoechst33342荧光染料和PI染色,流式细胞仪分析并分选SP细胞,并再次经流式细胞仪分析验证.采用免疫荧光及RT-PCR分析SP细胞ABCG2,CD133的表达情况.MTT法分析SP细胞及NSP细胞的增殖能力.结果 人神经母细胞瘤细胞系SH-SY5Y及WN中存在SP细胞,其比例在0.2％～1.3％左右,维拉帕米阻断后减少为0.1％～0.5％左右.免疫荧光及RT-PCR分析表明SP细胞表达ABCG2及CD133.MTT证实SP细胞与NSP细胞的增殖能力有差异(P＜0.05).结论 人神经母细胞瘤细胞系细胞中的SP细胞具有肿瘤干细胞特性.

abstractsObjective To explore side population(SP) cells in human neuroblastoma cell lines SH-SY5Y,WN and to observe the expression of the universal stem cell markers ABCG2 and CD133.Methods Neuroblastoma cells were stained with Hoechst33342 and PI.The SP cells were analyzed and sorted by the fluorescence-activated cell sorter.Then the expression of ABCG2 and CD133 was analyzed by immunofluorescence and RT-PCR method.Proliferation of SP and non-SP cells was measured with MTT colorimetric assay.Results SP cells accounted for 0.2％～1.3％ of total human neuroblastoma cells in culture,which decreased to 0.1％ ～0.5％ after addition of verapamil.SP cells were shown to express ABCG2 and CD133 using immunofluorescence and RT-PCR method.SP and non-SP cells had disparity in cell growth as assessed by MTT colormetric assay (P ＜ 0.05).Conclusions SP cells in neuroblastoma cell lines had cancer stem cell-like properties.