Identifying tumor stem-like cells in mouse melanoma cell lines by analyzing the characteristics of side population cells.
Increasingly more evidence shows that TSCs possess the characteristics of stem-like cells. However, a link between stem cells and TSCs remains to be shown. We have sorted SP cells and non-SP cells from the B16F10 cell lines by FACS, and then studied their cellular biological characteristics by using a SFCM culture method, proliferative assay in vitro, clone formation assays in soft agar and normal media, tumorigenic assays in C57BL/6 mice, and resistance to chemotherapy assay in vitro, the quantitative detecting expression of ABCG2 and their CD phenotype analysis by a FCM. We detected 0.96% SP cells in the B16F10 cells and found that they had obvious differences in characteristics from non-SP cells. They possessed a marked capacity for self-renewal in soft agar and culture medium, strong tumorigenic potential in C57BL/6 mice, apparent resistance to vinblastin in vitro, upregulated ABCG2 expression, and a high expression of CD44+CD133+CD24+ phenotypes. We conclude that there were a few of SP cells that had the characteristics of tumor stem-like cells which may provide a useful tool and a readily accessible source for further study when specific TSCs markers are unknown.
Dou
Jun
J
Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, 87# Dingjiaqiao Rd., Nanjing 210009, Jiangsu, China. njdoujun@yahoo.com.cn
Wen
Ping
P
Hu
Weihua
W
Li
Yating
Y
Wu
Yun
Y
Liu
Chunsheng
C
Zhao
Fengshu
F
Hu
Kai
K
Wang
Jing
J
Jiang
Chuilian
C
He
Xiangfeng
X
Gu
Ning
N
D000071916:AC133 Antigen; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D051927:CD24 Antigen / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000378:metabolism; D008546:Melanoma, Experimental / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D010455:Peptides / Q000378:metabolism; D014747:Vinblastine / Q000494:pharmacology
D000071916:AC133 Antigen; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; C501412:Abcg2 protein, mouse; D015703:Antigens, CD; D051927:CD24 Antigen; D006023:Glycoproteins; D018960:Hyaluronan Receptors; D010455:Peptides; C000606024:Prom1 protein, mouse; D014747:Vinblastine