人卵巢癌细胞系干/祖细胞分离及其鉴定  

Isolatian and identification of cancer stem/progenitor cells from human ovarian cancer cell line

作　　者：王翠环[1] 杜涌瑞[2] 高燕[2] 顾欣[1] 赵惠丰[1] 李宏钊[1] 姚智[1] 邓为民[1] 

机构地区：[1]天津医科大学基础医学院免疫学教研室天津市细胞与分子免疫学重点实验室,天津300070 [2]天津市中心妇产科医院,天津300052

出　　处：《中华肿瘤防治杂志》2010年第6期412-416,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30670801);天津市科学技术委员会资助项目(06YFJMJC08300)

摘　　要：目的:自人卵巢癌细胞系SK-OV-3中分离干/祖细胞并进行鉴定。方法:采用无血清球形体形成法从SKOV-3中分离培养卵巢癌干/祖细胞;采用实时定量PCR和蛋折质印迹法测定球形体细胞干/祖细胞相关标志ABCG2、Oct-4、Nanog基因和蛋白的表达;流式细胞仪检测其耐药性;双层软琼脂检测其克隆形成能力;NOD/SCID小鼠检测其体内致瘤性。结果:球形体细胞表达干/祖细胞相关标志Oct-4、ABCG2、Nanog;对顺铂高耐药;在双层软琼脂上克隆形成率达(13.67±1.48)%;1 000个球形体形成细胞就能在NOD/SCID鼠中成瘤。结论:采用无血清培养基中球形体形成法从SKOV-3细胞系中可以分离出具有干/祖特性的卵巢癌细胞,可为今后研究卵巢癌的发生、发展、复发及其化疗药物筛选提供简便实用的体外模型。OBJECTIVE： To isolate and identify the ovarian cancer stem/progenitor cells from human ovarian cancer cell line SKOV-3.METHODS： To isolate ovarian cancer stem/progenitor cells by sphere-forming in serum-free culture medium and to identify the stemness of this kind of cell from following aspects： 1） to determine the expression of genes and proteins of the stem/progenitor cell-relevant markers ABCG2,Oct-4,Nanog by Real-time PCR and Western blot respectively;2） to detect the drug resistance against cisplatin by flow cytometry;3） to detect the clone-formation ability in double-layer soft agar;4） to evaluate the tumor-formation ability in the NOD/SCID mice.RESULTS： The spheroid cells had unique growth characteristics and expressed stem/progenitor cell-relevant markers ABCG2,Oct-4 and at high level;and they had a high resistance to cisplatin and cloning forming efficiency of（13.67±1.48）%.Furthermore xenograft tumor formed only by 1 000 such cells in NOD/SCID mice.CONCLUSION： The ovarian cancer stem/progenitor cells can be isolated by sphere-forming in serum-free culture medium from SKOV-3,and can be used as a potential tool for scanning the anti-cancer drugs in the treatment of human ovarian cancer isolation and identification.

关 键 词：卵巢癌 癌干/祖细胞 细胞系 分离鉴定 

分 类 号：R737.31[医药卫生—肿瘤]