卵巢癌腹水中肿瘤干/祖细胞的筛选及鉴定  

Isolation and Identification of Cancer Stem/Progenitor Cells from Ascites of Patients with Ovarian Cancer

作　　者：王翠环[1] 高燕[1] 贺晓英[1] 杜涌瑞[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科,300070

出　　处：《天津医药》2010年第4期303-305,354,共4页Tianjin Medical Journal

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

摘　　要：目的:从人卵巢癌腹水细胞中分离肿瘤干/祖细胞并进行鉴定。方法:采用无血清培养法从腹水中分离培养球形体细胞(ASC);用流式细胞仪检测腹水ASC和腹水细胞(AC)中SP(side population)亚群含量;采用PCR和Western blot方法测定两种细胞干/祖细胞相关标志物ABCG2、Oct-4、Nanog基因和蛋白的表达;NOD/SCID小鼠检测其体内致瘤性。结果:腹水球形体细胞和腹水细胞中SP亚群分别为0.9%,0.2%;腹水球形体细胞较腹水细胞高表达干/祖细胞相关标志物Oct-4、ABCG2、Nanog;1×104个球形体细胞就能在NOD/SCID鼠中成瘤,而1×106个腹水细胞也不能成瘤。结论:无血清培养法可以从卵巢癌患者腹水中筛选出肿瘤干/祖细胞。该结果可为今后研究卵巢癌的发生、复发机制及筛选其化疗药物提供简便实用的体外模型。Objective：The and identify the stem/progenitor cells from the ascites of patients with ovarian cancer. Methods：The ovarian cancer stem/progenitor cells were isolated by sphere-forming assay in serum-free culture medium ,and were identified from side populations（SP） by flow-cytometry. The expressions of genes and proteins of the stem/progenitor cell-relevant markers ABCG2,Oct-4 and Nanog in ascetic spheroid cells and ascetic cells were determined by PCR and Western blot respectively. The tumor-formation ability was evaluated in the NOD/SCID mice. Results：The ascitic spheroid cells had unique growth characteristics,and SP cells accounted for 0.9% and 0.2% in ascitic spheroid cells and ascitic cells respectively. The ascitic spheroid cells expressed higher stem/progenitor cell-relevant markers ABCG2,Oct-4 and Nanog than that of ascitic cells. Furthermore,xenograft tumor was formed only by 1×10^4 ascitic spheroid cells in NOD/SCID mice. Conclusion：The ovarian cancer stem/progenitor cells can be isolated by sphere-forming assay in serum-free culture medium from ascites and can be used as a potential tool for studying the pathogenesis and scanning the anti-cancer drugs in the treatment of human ovarian cancer.

关 键 词：卵巢肿瘤 腹水液 肿瘤干细胞 球形体 细胞 生物学标记 小鼠 SCID 

分 类 号：R737.310.5[医药卫生—肿瘤]