Isolation and in vitro characterization of CD133 + side population cells from laryngeal cancer cell line
10.3760/cma.j.issn.1673-0860.2011.09.012
- VernacularTitle:喉癌CD133+侧群细胞的分离及体外生物学特性分析
- Author:
Liang ZHOU
1
;
Chun-Ping WU
;
Ming XIE
;
Lei TAO
;
Ming ZHANG
;
Jie TIAN
Author Information
1. 复旦大学附属眼耳鼻喉科医院
- Keywords:
Laryngeal neoplasms;
Neoplastic stem cells;
Glycoproteins;
Peptides;
Antigens,CD;
Side-population cells
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2011;46(9):752-757
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate an approach enriching cancer stem cells (CSCs) more effectively from laryngeal cancer cell line. Methods CD133 + 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. Results CD133 + 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.Conclusions Compared 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.
