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This version published online on January 18, 2007
Endocrinology, doi:10.1210/en.2006-1553
A more recent version of this article appeared on April 1, 2007
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Submitted on November 20, 2006
Accepted on January 10, 2007

Characterization of side population in thyroid cancer cell lines: cancer stem-like cells are enriched partly but not exclusively

Norisato Mitsutake*, Atsuhiko Iwao, Kazuhiro Nagai, Hiroyuki Namba, Akira Ohtsuru, Vladimir Saenko, and Shunichi Yamashita

Department of Molecular Medicine, Department of International Health and Radiation Research, Atomic Bomb Disease Institute, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan; Transfusion Service, Takashi Nagai Memorial International Hibakusha Medical Center, Nagasaki University Hospital of Medicine and Dentistry, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan

* To whom correspondence should be addressed. E-mail: mitsu{at}nagasaki-u.ac.jp.

There is increasing evidence that cancers contain their own stem-like cells called cancer stem cells (CSCs). A small subset of cells, termed "side population (SP)", has been identified using flow cytometric analysis. The SP cells have the ability to exclude the DNA binding dye, Hoechst33342, and are highly enriched for stem cells in many kinds of normal tissues. Since CSCs are thought to be drug-resistant, SP cells in cancers might contain CSCs. We initially examined the presence of SP cells in several human thyroid cancer cell lines. A small percentage of SP cells were found in ARO (0.25%), FRO (0.1%), NPA (0.06%) and WRO (0.02%) cells, but not in TPC1 cells. After sorting, the SP cells generated both SP and non-SP cells in culture. The clonogenic ability of SP cells was significantly higher than that of non-SP cells. Moreover, the SP prevalence was dependent on cell density in culture, suggesting that SP cells preferentially survived at lower cell density. Microarray experiment revealed differential gene expression profile between SP and non-SP cells, and several genes related to stemness were up-regulated. However, non-SP population also contained cells that were tumorigenic in nude mice, and non-SP cells generated a small number of SP cells. These results suggest that cancer stem-like cells are partly, but not exclusively, enriched in SP population. Clarifying the key tumorigenic population might contribute to the establishment of a novel therapy for thyroid cancer.
Key words: side population • cancer stem cells • thyroid cancer




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