CD44^+CD24^(-/low)乳腺癌干细胞的流式分选及其肿瘤干细胞特性的初步鉴定  

Isolation and Preliminary Characterization of CD44^+CD24^(-/low) Human Breast Cancer Stem Cells

作　　者：王玲燕[1] 王斌[2] 马清华[2] 金泗虎[1] 高红伟[1] 李素波[1] 卞修武[2] 宫锋[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850 [2]第三军医大学西南医院病理研究所和西南癌症中心,重庆400038

出　　处：《生物技术通讯》2012年第1期56-59,共4页Letters in Biotechnology

摘　　要：目的:通过表面标志分选法富集乳腺癌干细胞,并初步鉴定其肿瘤干细胞特性。方法:采用流式细胞分选术从人乳腺癌细胞系MCF-7中分选CD44+CD24-/low乳腺癌干细胞,并进行干细胞比例分析;用免疫荧光法检测、比较分选获得的细胞和对照细胞的干性和分化标记物Oct-4、SOX-2、CK-18和α-SMA的表达状态。结果:分选获得的CD44+CD24-/low乳腺癌干细胞阳性比例达90%以上;免疫荧光检测结果显示,CD44+CD24-/low细胞亚群比non-CD44+CD24-/low细胞亚群高表达干细胞转录因子Oct-4、SOX-2,低表达分化因子CK-18、α-SMA;体外实验表明,CD44+CD24-/low细胞亚群具有更强的成球生长能力,并具有双向分化潜能。结论:CD44+CD24-/low表面标记物分选的方法可以富集高纯度的乳腺癌干细胞,且呈现干性因子Oct-4和SOX-2高表达。Objective： To enrich breast cancer stem cells through fluorescence-activated cell sorting and verify their biological characteristics of the cells.Methods： CD44＋CD24-/low subset cells were sorted and measured by flow cytometry,and the expressions of Oct-4,SOX-2,CK-18 and α-SMA were detected by immunostaining.Results：Flow cytometry analysis indicated that the purity of CD44＋CD24-/low subset cells exceed 90%.Immunostaining analysis suggested that CD44＋CD24-/low subset cells expressed higher levels of transcription factors Oct-4 and SOX-2,lower levels of differentiated factors CK-18 and α-SMA than non-CD44＋CD24-/low subset cells.In vitro,CD44＋CD24-/low subset cells were verified to possess the ability of forming tumor spheres and multipotency.Conclusion： CD44＋CD24-/low subset cells contained higher proportion of breast cancer stem cells and expressed higher levels of Oct-4 and SOX-2.

关 键 词：乳腺癌干细胞 流式细胞分选 细胞分化 细胞表面分子标记 

分 类 号：Q25[生物学—细胞生物学]