Isolation and in vivo tumorigenicity assay of CD133+ side population cells from laryngeal cancer cell line / 中华耳鼻咽喉头颈外科杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate a valuable strategy for further purifying cancer stem cells (CSCs) from laryngeal cancer cell line.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>