免疫磁珠法分离并鉴定前列腺癌类干细胞.docVIP

免疫磁珠法分离并鉴定前列腺癌类干细胞[基金项目] [基金项目] 教育部新教师基金项目（No. 200804871051），湖北省卫生厅青年人才基金（QJX2010－5） [第一作者] 秦亮，电话E-mail: ql_198572@163.com（稿件相关事宜烦请与第一作者联系） [通信作者] 廖晖，E-mail: liaohui@ 秦亮，陈安民，郭风劲，卞峰，杜宇，姚斌，廖晖（430030 湖北武汉，华中科技大学同济医学院附属同济医院骨科） [摘要] 目的 利用免疫磁珠法分选并鉴定人前列腺癌类干细胞。方法 利用免疫磁珠法从人前列腺癌细胞株PC-3中分选出CD133+/CD44+细胞，通过免疫组化、流式细胞术、CCK8法、平板克隆形成实验以及体外裸鼠移植瘤实验鉴定其表面标志物表达情况及其肿瘤干细胞特性。结果 经免疫磁珠分选所得CD133+/CD44+细胞数比例为0.50%；免疫组化结果显示其CD44/CD133/integinα2β1均呈高表达；流式细胞分析显示其CD44/CD133双阳性率达52.28%，其同时表达integinα2β1阳性的细胞比例为59.16%；其细胞增殖率和克隆形成率高于PC-3细胞；以低密度（1×1 03）皮下注射时，其裸鼠体内成瘤率明显高于PC-3细胞。结论 利用免疫磁珠分选法可以从人前列腺癌细胞株PC-3中高效的分选出具有肿瘤干细胞生物学特性的前列腺癌类干细胞。 [关键词] 肿瘤干细胞；前列腺癌；免疫磁珠分选 The isolation and identification of the prostate cancer stem cells by magnetic active cell sorting. Qin Liang, Chen An-min, Guo Fengjing, Bian Feng, Du Yu, Yao Bin, Liao Hui. (Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China) [ABSTRACT] Objective To isolate, culture and identify the prostate cancer stem cells (CSCs) by magnetic active cell sorting (MACS). Methods CD133+/CD44+ cells were selected from PC-3 human prostatic carcinoma cells lines by MACS, and they were identified as prostate cancer stem-like cells by immunohistochemistry, flow-cytometric analysis, CCK8, colony-forming assay and tumorigenicity assay in vivo. Results CD133+/CD44+ cells selected from PC-3 cell lines by MACS showed that the proportion of CD133+/CD44+ cells was 52.28%, and the proportion of integinα2β1+ cells was 59.16% by flow-cytometric analysis; the selected cells proliferating potential and clony-forming rate were higher than PC-3 cell lines in vitro; and their tumorigenicity rate were significantly higher than PC-3 cell lines in vivo when seeded at a low density(1×103). Conclusion MACS was a simple and effective method to gain prostatic carcinoma stem cells from PC-3 human prostatic carcinoma cell lines. [Key words] Cancer stem cells; Prostate cancer; Magnetic active cell