天津医药

• 实验研究 •    

卵巢癌腹水中肿瘤干/祖细胞的筛选及鉴定

王翠环1,高燕2,贺晓英3,杜涌瑞4,姚智4,邓为民5   

  1. 1. 天津医科大学基础医学院免疫教研室
    2. 天津市中心妇产医院
    3. 天津医科大学
    4.
    5. 天津医科大学免疫教研室
  • 收稿日期:2009-11-30 修回日期:2009-12-10 出版日期:2010-04-15 发布日期:2010-04-15
  • 通讯作者: 王翠环

The isolation and identification of cancer stem/progenitor cells from the ascites of ovarian cancer Patients

  • Received:2009-11-30 Revised:2009-12-10 Published:2010-04-15 Online:2010-04-15

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摘要： 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：采用无血清特殊培养法从腹水中分离培养球形体细胞；用流式细胞仪检测腹水球形体细胞（ascitic spheroid cells，ASC）和腹水细胞（ascitic cells，AC）中SP(side population) 亚群含量；采用PCR和Western blot方法测定两种细胞干/祖细胞相关标记物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/祖细胞相关标记物Oct-4、ABCG2、Nanog；1×104个球形体形成细胞就能在NOD/SCID鼠中成瘤，而腹水细胞1×106个细胞也不能成瘤。结论：无血清悬浮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。

关键词: 卵巢癌, 肿瘤干/祖细胞, 腹水, 分离鉴定

Abstract: The isolation and identification of cancer stem/progenitor cells from the ascites of ovarian cancer Patients WANG Cuihuan, GAO Yan, HE Xiaoying, DU Yongrui, YAO Zhi, DENG Weiming△Department of immunology, Tianjin Medical University, Tianjin 300070, China ? Abstract Objective: To isolate and identify the stem/progenitor cells from the ascites of ovarian cancer patients. 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: side population were detected by the flow-cytometry; to determine the expression of genes and proteins of the stem/progenitor cell-relevant markers ABCG2, Oct-4, Nanog in ascetic spheroid cells and ascetic cells by PCR and Western blot respectively; to evaluate the tumor-formation ability 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 ascitic cells; Furthermore，xenograft tumor formed only by 1×104 ascitic spheroid cells in NOD/SCID mice. Conclusions: The ovarian cancer stem/progenitor cells can be isolated by sphere-forming 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. Key words ovarian cancer tumor stem/ progenitor cells isolation identification

Key words: ovarian cancer, tumor stem/ progenitor cells, isolation, identification