[Isolation and in vivo tumorigenicity assay of CD133+ side population cells from laryngeal cancer cell line].
OBJECTIVE
To investigate a valuable strategy for further purifying cancer stem cells (CSCs) from laryngeal cancer cell line.


METHODS
CD133+ side population (SP) and CD133-SP cells were detected and isolated from laryngeal cancer Hep-2 cell line with SP discrimination and CD133 surface marker, assisted by fluorescence activated cell sorting technology. Freshly sorted CD133+SP and CD133-SP cells were xenografted into the subcutaneous space of the right axillary fossa of NOD/SCID mice and tumorigenic capacity of the cells from two subgroups were examine. Cell cycle distributions of the two cell populations were detected.


RESULTS
CD133+SP and CD133-SP cells accounted for (0.30±0.12)% and (17.52±1.59)% in Hep-2 cell line, respectively. CD133+SP cells formed tumor nodules in 15 of 16 mice and CD133-SP cells in 7 of 16 mice (Fisher's exact test, P<0.05). The mean weight of CD133+SP tumor nodules was (0.36±0.15)g and that of CD133-SP tumor nodules was (0.08±0.04) g. The difference was significant (t=4.64, P<0.01). Cell cycle analysis revealed similar cycle distributions between the two subgroups.


CONCLUSIONS
CD133+SP cells harbored much more cancer stem-like tumorigenic potential in NOD/SCID mice than CD133-SP cells. The combination of SP discrimination and surface marker selection helped to purify CSCs further from laryngeal cancer cell line.
Department of Otorhinolaryngology Head and Neck Surgery, Fudan University Affiliated Eye, Ear, Nose and Throat Hospital, Shanghai 200031, China.
Chun-ping
CP
Wu
Liang
L
Zhou
Ming
M
Xie
Lei
L
Tao
Ming
M
Zhang
Jie
J
Tian
D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D007822:Laryngeal Neoplasms; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000166:cytology*; D010455:Peptides / Q000032:analysis; D058985:Side-Population Cells / Q000166:cytology*
D000071916:AC133 Antigen; D015703:Antigens, CD; D014408:Biomarkers, Tumor; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse