中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2013年
5期
325-329
,共5页
神经母细胞瘤%侧群细胞%肿瘤干细胞%ABCG2%CD133
神經母細胞瘤%側群細胞%腫瘤榦細胞%ABCG2%CD133
신경모세포류%측군세포%종류간세포%ABCG2%CD133
Neuroblastoma%Side population cells%Neoplastic stem cells%ABCG2%CD133
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.
目的 檢測人神經母細胞瘤細胞繫SH-SY5Y及WN中是否存在側群(side population,SP)細胞,併研究該SP細胞是否具有榦細胞某些生物學特性.方法 經Hoechst33342熒光染料和PI染色,流式細胞儀分析併分選SP細胞,併再次經流式細胞儀分析驗證.採用免疫熒光及RT-PCR分析SP細胞ABCG2,CD133的錶達情況.MTT法分析SP細胞及NSP細胞的增殖能力.結果 人神經母細胞瘤細胞繫SH-SY5Y及WN中存在SP細胞,其比例在0.2％～1.3％左右,維拉帕米阻斷後減少為0.1％～0.5％左右.免疫熒光及RT-PCR分析錶明SP細胞錶達ABCG2及CD133.MTT證實SP細胞與NSP細胞的增殖能力有差異(P＜0.05).結論 人神經母細胞瘤細胞繫細胞中的SP細胞具有腫瘤榦細胞特性.
목적 검측인신경모세포류세포계SH-SY5Y급WN중시부존재측군(side population,SP)세포,병연구해SP세포시부구유간세포모사생물학특성.방법 경Hoechst33342형광염료화PI염색,류식세포의분석병분선SP세포,병재차경류식세포의분석험증.채용면역형광급RT-PCR분석SP세포ABCG2,CD133적표체정황.MTT법분석SP세포급NSP세포적증식능력.결과 인신경모세포류세포계SH-SY5Y급WN중존재SP세포,기비례재0.2％～1.3％좌우,유랍파미조단후감소위0.1％～0.5％좌우.면역형광급RT-PCR분석표명SP세포표체ABCG2급CD133.MTT증실SP세포여NSP세포적증식능력유차이(P＜0.05).결론 인신경모세포류세포계세포중적SP세포구유종류간세포특성.
Objective 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.
