摘要
背景与目的:数量上的稀少和不断分化的能力阻碍了肿瘤干细胞的分离和纯化,现有的筛选方法只是富集了异质性的肿瘤干细胞群,无法取得更为原始的、接近源头的肿瘤干细胞。本文以肿瘤干细胞假说为理论依据,探讨悬浮培养联合化疗药物分离、培养、纯化及鉴定小鼠乳腺癌干细胞的可行性研究。方法:无血清悬浮培养小鼠乳腺癌TM40D细胞,流式细胞仪检测CD44+CD24-细胞含量。选取CD44+CD24-含量最多代数的细胞,加入不同血浆峰浓度(peak plasma concentrations,PPC)的联合化疗药物紫杉醇和表柔比星,作用24 h后继续无血清培养,Annexin-V FITC/PI流式细胞仪法检测细胞凋亡。将筛选得到的细胞以不同的数量接种到BALB/C小鼠皮下,观察致瘤能力。结果:无血清悬浮培养第10代的细胞中CD44+CD24-比例最高(77%)。接种0.35 PPC组100个细胞即可见肿瘤生成。结论:小鼠乳腺癌TM40D细胞中存在肿瘤干细胞,悬浮培养联合化疗药物筛选乳腺癌干细胞在方法上是可行的。
Background and purpose： The scarcity in quantity and continuous differentiation ability severely compromise the isolation and purification of cancer stem cells （ CSCs）. The existing sorting methods can only enrich CSCs as a het-erogeneous population. However, more primitive CSCs closer to the source still have not been identified. Based on the CSC hypothesis, we hereby explored the possibility of isolation, culture, purification and identification of murine breast cancer stem cells in suspension culture combined with anticancer regimens. Methods： TM40D murine breast cancer cells were cul- tured in serum-free medium. The expression of CD44^＋CD24^- was measured by flow cytometry. Cells from the passage of TM40D cells with the highest expression of CD44^＋CD24^- were treated in combination with anticancer agents pacilitaxel and epirubicin at different peak plasma concentrations （PPC） for 24 hours, and then maintained under suspension culture. The rate of apoptosis was examined by flow cytometry with Annexin-V FITC/PI double staining method. Selected cells in different amounts were injected subcutaneously into BALB/C mice to observe tumor formation. Results： Cells of passage 10 in suspension culture had the highest percentage of CD44^＋CD24^-（ about 77 percent）. As few as 100 cells in 0.35 PPC could generate tumors in BALB/C mice. Conclusions： TM40D cell line contains breast cancer stem cells. This study suggested that suspension culture combined with anticancer regimens is a feasible approach for screening tumor stem cells.
出处
《中国癌症杂志》
CAS
CSCD
2008年第3期172-175,共4页
China Oncology
关键词
肿瘤干细胞
悬浮培养
化疗药物
乳腺癌
移植瘤
cancer stem cells
suspension culture
chemotherapeutic agents
breast cancer
xenograft
