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CD44 is of Functional Importance for Colorectal Cancer Stem Cells

Authors' Affiliations: ¹ Joint Laboratory of Apoptosis and Cancer Biology, the State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, and College of Life Sciences, Nankai University, ² Graduate University of the Chinese Academy of Sciences, ³ Laboratory of Molecular Oncology, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing, China, ⁴ Cancer Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China, and ⁵ INSERM U756, Faculté de Pharmacie, Université Paris-Sud, Chatenay-Malabry, France

Requests for reprints: Quan Chen, The Laboratory of Apoptosis and Cancer Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, P.R. China. Phone: 86-10-6480-7321; Fax: 86-10-6480-7321; E-mail: chenq{at}ioz.ac.cn or Youyong Lu, Laboratory of Molecular Oncology, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing 100142, P.R. China. Phone: 86-10-8819-6765; E-mail: yongylu{at}public.bta.net.cn.

Purpose: Both CD44 and CD133 were reported as putative markers for isolating colorectal cancer stem cells (CSC). It remains to be resolved if both of these markers are of functional importance for colorectal CSC.

Experimental Design: The expression of CD44 and CD133 in normal colonic tissues and primary colorectal cancer was assessed by immunohistochemistry in a series of 60 patients on tissue microarray sections. Both in vitro clonogenic and in vivo tumorigenic assay were applied to measure CSC activities from the cells isolated from patients. Lentiviral RNA interference was used to stably knock down CD44 or CD133 in colorectal cancer cells from patients.

Results: We found that CD44⁺ cells displayed clustered growth and they did not colocalize with CD133⁺ cells within colorectal cancer. As few as 100 CD44⁺ cells from a patients' tumor initiated a xenograft tumor in vivo. A single CD44⁺ cell from a tumor could form a sphere in vitro which has characteristic stem cell properties and was able to generate a xenograft tumor resembling the properties of the primary tumor. Knockdown of CD44, but not CD133, strongly prevented clonal formation and inhibited tumorigenicity in xenograft model.

Conclusions: These results indicate that CD44 is a robust marker and is of functional importance for colorectal CSC for cancer initiation.

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