Isolation and in vitro characterization of CD133(+) side population cells from laryngeal cancer cell line.
- Author:
Chun-ping WU
1
;
Liang ZHOU
;
Ming XIE
;
Lei TAO
;
Ming ZHANG
;
Jie TIAN
Author Information
- Publication Type:Journal Article
- MeSH: AC133 Antigen; Antigens, CD; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cell Separation; Glycoproteins; Humans; Laryngeal Neoplasms; pathology; Neoplastic Stem Cells; cytology; Peptides; Side-Population Cells; cytology
- From: Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(9):752-757
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate an approach enriching cancer stem cells (CSCs) more effectively from laryngeal cancer cell line.
METHODSCD133(+)SP and CD133(-)SP subpopulation was detected and isolated from Hep-2 cell line using Hoechst33342 dye and phycoerythrin (PE)-conjugated CD133 monoclonal antibody assisted by fluorescence activated cell sorting technology. Sorted CD133(+)SP and CD133(-)SP cells were compared in CSCs-related assays including proliferation, differentiation, spheroid formation and drug sensitivity.
RESULTSCD133(+)SP cells accounted for a very small fraction of (0.30 ± 0.12)% in Hep-2 cell line, far less than the proportion of CD133(+) subgroup and side population subgroup, which were (3.15 ± 0.83)% and (17.1 ± 2.0)% respectively. Intriguingly, CD133(+)SP cells proliferated much faster than CD133(-)SP cells in RPMI1640 and gave rise to CD133(-)SP cells and other heterogeneous cells that formed the bulk of the tumor. In contrast, CD133(-)SP cells were not able to differentiate into CD133(+)SP cells. In serum-free medium CD133(+)SP cells grew as spherical clusters and remained floating. In addition, CD133(+)SP cells manifested the marked resistance to chemotherapy than CD133(-)SP cells.
CONCLUSIONSCompared with CD133(-)SP cells, CD133(+)SP subpopulation exhibited extraordinary cancer stem-like properties, were enriched for cancer stem cells more effectively and might serve as an ideal putative candidate for CSCs research in laryngeal cancer.
