人前列腺癌PC-3细胞中肿瘤干细胞的分离及鉴定  

Isolation and identification of prostate cancer stem cells from PC-3 cells

作　　者：周耀军[1] 许贤林[1] 徐仁芳[1] 崔笠[1] 夏炜[1] 曹赟杰[1] 范敏[1] 何小舟[1] 

机构地区：[1]苏州大学附属第三医院(常州市第一人民医院)泌尿外科,常州213003

出　　处：《中华实验外科杂志》2015年第9期2207-2210,共4页Chinese Journal of Experimental Surgery

基　　金：江苏省自然科学基金资助项目（BK20141161）

摘　　要：目的 动态观察PC-3细胞克隆在形态上的差异,检测CD44在不同克隆中的表达,比较不同克隆细胞在裸鼠体内的成瘤能力,并鉴定含前列腺癌干细胞的克隆.方法 克隆形成实验观察PC-3细胞克隆的形态并分类,计算克隆形成率及每种克隆的比例,运用免疫荧光技术（IF）检测不同克隆CD44的表达.分离不同形态的克隆,检测其自我更新能力.将不同形态的克隆细胞分别接种于裸鼠皮下,观察各组成瘤能力.结果 PC-3细胞可形成3种克隆形态,即Holoclone、Meroclone及Paraclone.克隆形成率为（9.30±1.61）％（n=5）,Holoclone、Meroclone和Paraclone 3种克隆的比例分别为10.0％、48.3％和41.7％.免疫荧光检测结果显示,CD44在Holoclone中的表达明显强于Meroclone及Paraclone.Holoclone具有自我更新能力,在裸鼠体内具有较强的成瘤能力.结论 PC-3细胞的克隆在形态和分化程度上存在差异,Holoclone富含前列腺癌干细胞.Objective To dynamically observe the morphology of PC-3 human prostate cancer cell colonies,detect the expression of CD44 and then compare the tumorigenic ability of different clones in order to identify the cancer stem cell clones.Methods Colony formation assay was used to observe and classify the colonies derived from PC-3 human prostate cancer cells,then cloning efficiency and proportion of each colony were calculated.The expression of CD44 was detected by immunofluorescence microscopy.Furthermore,different clones were isolated and cultured,and their self-renewal property was analyzed.Cells of different clones were inoculated into nude mice subcutaneously and their tumorigenic ability was determined.Results Colonies derived from PC-3 human prostate cancer cells had three types,such as Holoclone,Meroclone and Paraclone.Cloning efficiency was 9.3％,and the proportion of Holoclone,Meroclone and Paraclone colonies was 10.0％,48.3％ and 41.7％ respectively.The results of immunofluorescence microscopy showed the signal of CD44 was dramaticlly stronger in the Holoclone colonies than in the Meroclone and Paraclone.PC-3 cells of Holoclone clones possessed strong ability of self-renewal and in vivo tumorigenicity in the nude mice.Conclusion PC-3 cell colonies varied in morphology and difrerentiation,and the Holoclone colonies might be comprised of cancer stem cells.

关 键 词：PC-3细胞 前列腺癌 肿瘤干细胞 克隆 

分 类 号：R730.1[医药卫生—肿瘤]