摘要
目的:检测原代培养的乳腺癌细胞中乙醛脱氢酶1阳性(aldehyde dehydrogenase 1-positive,ALDH1+)和CD44+CD24-/low标志的乳腺癌干细胞的生物学特性差异,并探讨同时表达这2种干细胞标志的肿瘤干细胞亚群(即ALDH1+CD44+CD24-/low细胞)的生物学特性。方法 :采用FCM技术从新鲜乳腺癌组织的原代培养肿瘤细胞中分选出CD44+CD24-/low、ALDH1+和ALDH1+CD44+CD24-/low细胞亚群,分析各分选细胞亚群占总肿瘤细胞的比例。采用免疫荧光法证实原代培养的肿瘤细胞中存在各分选细胞;然后采用克隆形成实验、MTT法和Transwell侵袭实验观察各组分选细胞的自我更新、增殖和侵袭能力,并通过裸鼠体内成瘤实验观察各组分选细胞在裸鼠体内的致瘤能力。结果 :CD44+CD24-/low细胞占总肿瘤细胞的比例为7.2%,ALDH1+细胞占4.6%,而ALDH1+CD44+CD24-/low细胞占1.5%。3组细胞的自我更新、增殖、侵袭及致瘤能力由强至弱依次为ALDH1+CD44+CD24-/low、ALDH1+和CD44+CD24-/low细胞,组间差异均有统计学意义(P值均<0.05)。结论:表达CD44+CD24-/low、ALDH1+和ALDH1+CD44+CD24-/low标志的3组乳腺癌干细胞亚群在自我更新、体外增殖和侵袭能力以及体内致瘤能力等生物学特性上均存在明显差异,其中以ALDH1+CD44+CD24-/low亚群最强,提示ALDH1+CD44+CD24-/low可作为特异性更强的乳腺癌干细胞标志。
Objective： To explore the differences in biological characteristics among aldehyde dehydrogenase 1-positive （ALDH1＋） and CD44^＋ CD24-/low-marked breast cancer stem cells from primary cultured breast cancer cells, and to investigate the biological properties of ALDH1^＋CD44^＋CD24^-/low-marked stem cell subpopulations.Methods： FCM was used to isolate CD44^＋CD24-/low, ALDH1^＋ and ALDH1^＋ CD44^＋CD24-/low stem cell subpopulations from the primary cultured breast cancer cells derived from fresh human breast cancer tissues, and the proportions of these three stem cell subpopulations were analyzed. The three isolated stem cell subpopulations were identified by immunofluorescence method, and their capacities of self-renewal, proliferation and invasion were analyzed by clone formation, MTT method and Transwell invasion assay, as well as the in vivo tumorigenicity abilities of the three stem cell subpopulations in nude mice were examined.Results： The CD44^＋CD24-/low, ALDH1^＋ and ALDH1^＋CD44^＋CD24-/low stem cell subpopulations were isolated from human breast cancer cells, and accounted for 7.2%, 4.6% and 1.5% of total tumor cells, respectively. According to the capacities of self-renewal, proliferation, invasion and tumorigenesis in order of increasing ability, these three breast cancer stem cell subpopulations were listed sequentially as ALDH1^＋CD44^＋CD24-/low, ALDH^＋ and CD44^＋CD24-/low, there were statistical significances among the three cell subpopulations （all P 〈 0.05）.Conclusion： There are significant differences in abilities of self-renewal, in vitro proliferation and invasion as well as in vivo tumorigenesis among CD44^＋CD24-/low, ALDH1^＋ and ALDH1^＋CD44^＋CD24-/low breast stem cell subpopulations, in which ALDH1^＋CD44^＋CD24-/low stem cell subpopulation has the strongest abilities, indicating that ALDH1^＋CD44^＋CD24-/low can be considered as a more specific marker for breast cancer stem cells.
出处
《肿瘤》
CAS
CSCD
北大核心
2015年第9期943-951,960,共10页
Tumor
基金
湖北省卫生厅科研重点项目(编号:JX4A07)~~
关键词
乳腺肿瘤
乙醛脱氢酶1
肿瘤干细胞
流式细胞术
小鼠
裸
Breast neoplasms
Aldehyde dehydrogenase 1
Neoplastic stem cells
Flowcytometry
Mice, nude
