Identification of a stem-like cell population by exposing metastatic breast cancer cell lines to repetitive cycles of hypoxia and reoxygenation.
Identification of a stem-like cell population by exposing metastatic breast cancer cell lines to repetitive cycles of hypoxia and reoxygenation.
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DOI：
10.1186/bcr2773
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Chen EI
中科院分区：
文献类型：
--
作者：
Louie E;Nik S;Chen JS;Schmidt M;Song B;Pacson C;Chen XF;Park S;Ju J;Chen EI
实体瘤不规则的血管系统造成缺氧区，其特点是缺氧和再氧循环周期。积累的证据表明，慢性和反复暴露于缺氧和再氧化似乎对肿瘤生长有利。尽管肿瘤缺氧耐受性的发展预示着不良预后，但促进缺氧耐受性的机制仍有待阐明。最近的研究描述了肿瘤内的癌症干细胞（CSC）亚群，它们具有干细胞样特性，如自我更新和分化成多种细胞类型的能力。肿瘤干细胞理论表明，CSCs作为一个独特的群体存在于肿瘤中，并通过产生新的肿瘤引起复发和转移。由于缺氧被认为是促进肿瘤侵袭性生长的关键生态位因素之一，我们假设缺氧/再氧化的重复循环也在乳腺CSCs的富集中发挥作用。以两种转移性人乳腺癌细胞系MDA-MB 231和BCM2为研究对象，优化了缺氧和复氧循环的条件。通过三色流式细胞术检测CD44、CD24、ESA和E-cadherin的表达，分析循环缺氧选择亚群中CSCs的百分比。用菌落形成试验来评估这个亚种群自我更新的能力。进行限制性稀释试验以评估该亚群的肿瘤启动和转移能力。通过表达EMT相关标记物和EMT相关microrna来检测EMT的诱导。利用优化的缺氧和再氧方案，我们从人类乳腺癌细胞系中鉴定出一种新的循环缺氧选择亚群，并证明了干细胞样乳腺癌细胞亚群可以通过重复缺氧/再氧循环而无需基因操作来扩增。我们还发现，来自这个新亚群的细胞很容易形成集落，在免疫缺陷小鼠中具有高度的致瘤性，并表现出干细胞样和EMT表型。这些结果为新建立的缺氧/复氧培养系统研究肿瘤微环境中生态位因子对乳腺癌细胞系CSCs的调控以及制定根除乳腺CSCs的差异化靶向策略提供了依据。
The irregular vasculature of solid tumors creates hypoxic regions, which are characterized by cyclic periods of hypoxia and reoxygenation. Accumulated evidence suggests that chronic and repetitive exposure to hypoxia and reoxygenation seem to provide an advantage to tumor growth. Although the development of hypoxia tolerance in tumors predicts poor prognosis, mechanisms contributing to hypoxia tolerance remain to be elucidated. Recent studies have described a subpopulation of cancer stem cells (CSC) within tumors, which have stem-like properties such as self-renewal and the ability to differentiate into multiple cell types. The cancer stem cell theory suggests CSCs persist in tumors as a distinct population and cause relapse and metastasis by giving rise to new tumors. Since hypoxia is considered to be one of the critical niche factors to promote invasive growth of tumors, we hypothesize that repetitive cycles of hypoxia/reoxygenation also play a role in the enrichment of breast CSCs. Two metastatic human breast cancer cell lines (MDA-MB 231 and BCM2) were used to optimize the conditions of hypoxia and reoxygenation cycles. The percentage of CSCs in the cycling hypoxia selected subpopulation was analyzed based on the CD44, CD24, ESA, and E-cadherin expression by three-color flow cytometry. Colony formation assays were used to assess the ability of this subpopulation to self-renew. Limiting dilution assays were performed to evaluate the tumor-initiating and metastatic ability of this subpopulation. Induction of EMT was examined by the expression of EMT-associated markers and EMT-associated microRNAs. Using an optimized hypoxia and reoxygenation regimen, we identified a novel cycling hypoxia-selected subpopulation from human breast cancer cell lines and demonstrated that a stem-like breast cancer cell subpopulation could be expanded through repetitive hypoxia/reoxygenation cycles without genetic manipulation. We also found that cells derived from this novel subpopulation form colonies readily, are highly tumorigenic in immune-deficient mice, and exhibit both stem-like and EMT phenotypes. These results provide the validity to the newly developed hypoxia/reoxygenation culture system for examining the regulation of CSCs in breast cancer cell lines by niche factors in the tumor microenvironment and developing differential targeting strategies to eradicate breast CSCs.
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影响因子：
7.4
作者：
Fillmore, Christine M.;Kuperwasser, Charlotte
通讯作者：
Kuperwasser, Charlotte
DOI：
10.4161/cc.8.20.9701
发表时间：
2009-10-15
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Heddleston JM;Li Z;McLendon RE;Hjelmeland AB;Rich JN
通讯作者：
Rich JN
影响因子：
4
作者：
Bolós, V;Peinado, H;Cano, A
通讯作者：
Cano, A
影响因子：
3.7
作者：
Javle, M. M.;Gibbs, J. F.;Khoury, T.
通讯作者：
Khoury, T.
影响因子：
7.3
作者：
Lang, S. H.;Frame, F. M.;Collins, A. T.
通讯作者：
Collins, A. T.