外阴鳞状细胞癌肿瘤干细胞的分选和鉴定

Separation and characterization of cancer stem like cells in vulvar squamous cell carcinoma

目的:分离外阴鳞状细胞癌A-431细胞系中CD44^+的细胞亚群,鉴定其具有肿瘤干细胞特性。方法:采用流式细胞术分选出CD44免疫荧光抗体标记的CD44^+细胞亚群,检测CD133、CD34蛋白在CD44^+和CD44^-中表达的差异,检测CD44^+及CD44^-细胞亚群的细胞周期。结果:A-431细胞系中CD44^+细胞亚群约占4%,胚胎干细胞因子CD133、CD34在CD44^+和CD44^-的VSCC中表达存在显著性差异,主要在细胞膜上表达,CD44^+的肿瘤细胞亚群97.69%处于G_1期,CD44^-的肿瘤细胞亚群61.12%处于S期。结论:在A-431细胞系中CD44可以作为分选肿瘤干细胞的特异性表面标志物,CD133、CD34在CD44^+的肿瘤细胞高表达,可能成为分选外阴鳞癌肿瘤干细胞的特异性表面标志物,CD44^+肿瘤细胞具有典型的干细胞增殖特点。

Objective：To isolate the cell subsets of CD44＋ from A-431 cell line,to prove that it has the characteristics of cancer stem cells.Methods：To isolate the cell subsets of CD44＋ by flow cytometry.To detect the expression of CD133,CD34 protein,comparing protein expression in the CD44＋ and CD44- cell population.To detect the cell cycle of the CD44＋ cell population and CD44- cell populations.Results：Cell subsets of CD44＋ in A-431 cells accounted for about 4%.CD44＋ cells differentially expressed the CD133,CD34 at protein levels,mainly expressed on the cell membrane.97.69% CD44＋ cancer cells lied in G1 period,61.12% CD44-cancer cells lied in S period.Conclusion：CD44 can become specificity surface marker for isolating cancer stem cells in A-431.The expression of CD133 and CD34 is higher in CD44＋ cells,and CD133,CD34 may become specificity surface markers for isolating CSC in VSCC.CD44＋ cancer cells have typical characteristics of stem cells proliferation.