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Blood, 1 December 2006, Vol. 108, No. 12, pp. 3906-3912.
Prepublished online as a Blood First Edition Paper on August 15, 2006; DOI 10.1182/blood-2006-05-024687.
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Submitted May 22, 2006
Accepted July 18, 2006
Characterization of stem cell like cancer cells
in immune competent mice
Jorg A Kruger, Charles D Kaplan, Yunping Luo, He Zhou, Dorothy Markowitz, Rong Xiang, and Ralph A. Reisfeld*
The Scripps Research Institute, Department of Immunology, La Jolla, California, USA
* Corresponding author; email: reisfeld{at}scripps.edu.
Recently, the cancer stem cell hypothesis has gained significant recognition as the descriptor of tumorigenesis. While previous studies relied on transplanting human or rat tumor cells into immune-compromised mice, our study utilized the Hoechst 33342 dye-based Side Population (SP) technique to isolate and transplant stem cell like cancer cells (SCLCC) from the 4T1 and NXS2 murine carcinoma cell lines into the immune-competent microenvironment of syngeneic mice. 4T1 cells displayed a SP of 2% with a Sca-1highc-Kit-CD45- phenotype, while NXS2 cells contained a SP of 0.2% with a Sca-1highCD24highc-Kit-CD45-GD2high phenotype. RT-PCR further revealed up-regulation in SP cells of ABCG2, Sca-1, Wnt-1 and TGF- 2. Additionally, 4T1 and NXS2 SP cells exhibited increased resistance to chemotherapy, and 4T1 SP cells also demonstrated an increased ability to efflux doxorubicin, which correlated with a selective increase in the percentage of SP cells found in the tumors of doxorubicin treated mice. Most importantly, SP cells demonstrated a markedly higher repopulation and tumorigenic potential in vivo, which correlated with an increased number of cells in the SP compartment of SP-derived tumors. Taken together, these results demonstrate that we successfully characterized SCLCCs from two murine carcinoma cell lines in the immune-competent microenvironment of syngeneic mice.
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