Phenotypic characterization of human colorectal cancer stem cells
Phenotypic characterization of human colorectal cancer stem cells
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DOI：
10.1073/pnas.0703478104
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发表时间：
2007-06-12
影响因子：
11.1
通讯作者：
Clarke, Michael F.
中科院分区：
文献类型：
--
作者：
Dalerba, Piero;Dylla, Scott J.;Clarke, Michael F.
最近的观察表明，在几种类型的人类癌症中，每个肿瘤中只有癌细胞的表型子集能够启动肿瘤生长。这种癌细胞的功能亚群在操作上被定义为“癌症干细胞”（CSC）亚群。在这里，我们开发了一个CSC模型，用于研究人类结直肠癌（CRC）。将实体CRC组织（从手术标本收集的原发组织或在非肥胖糖尿病/严重联合免疫缺陷（NOD/SCID）小鼠中建立的异种移植物）分解成单细胞悬液并通过流式细胞术进行分析。选择在上皮癌细胞中显示肿瘤内异质表达的表面标志物用于细胞分选和致瘤性实验。纯化单个表型癌细胞亚群，并通过注射NOD/SCID小鼠研究其肿瘤起始特性。我们的观察结果表明，在6/6的人CRC试验中，免疫缺陷小鼠体内移植的能力仅限于少数上皮细胞粘附分子（EpCAM）（高）/CD 44（+）上皮细胞亚群。起源于EPCAM（高）/CD 44（+）细胞的肿瘤保持分化表型，并再现其亲代病变的完整形态学和表型异质性。EpCAM（高）/CD 44（+）细胞的表面分子库的分析导致鉴定出CD 166作为额外的差异表达标记，其可用于在三个测试的CRC中的三个中的CSC分离。这些结果验证了人CRC中的干细胞工作模型，并为CRC干细胞分离提供了高度稳健的表面标志物谱。
Recent observations indicate that, in several types of human cancer, only a phenotypic subset of cancer cells within each tumor is capable of initiating tumor growth. This functional subset of cancer cells is operationally defined as the "cancer stem cell" (CSC) subset. Here we developed a CSC model for the study of human colorectal cancer (CRC). Solid CRC tissues, either primary tissues collected from surgical specimens or xenografts established in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice, were disaggregated into single-cell suspensions and analyzed by flow cytometry. Surface markers that displayed intratumor heterogeneous expression among epithelial cancer cells were selected for cell sorting and tumorigenicity experiments. Individual phenotypic cancer cell subsets were purified, and their tumor-initiating properties were investigated by injection in NOD/SCID mice. Our observations indicate that, in six of six human CRC tested, the ability to engraft in vivo in immunodeficient mice was restricted to a minority subpopulation of epithelial cell adhesion molecule (EpCAM)(high)/CD44(+) epithelial cells. Tumors originated from EPCAM(high)/CD44(+) cells maintained a differentiated phenotype and reproduced the full morphologic and phenotypic heterogeneity of their parental lesions. Analysis of the surface molecule repertoire of EpCAM(high)/CD44(+) cells led to the identification of CD166 as an additional differentially expressed marker, useful for CSC isolation in three of three CRC tested. These results validate the stem cell working model in human CRC and provide a highly robust surface marker profile for CRC stem cell isolation.