In vivo investigation of CD133 as a putative marker of cancer stem cells in Hep-2 cell line.
BACKGROUND
Mounting evidence suggests that most tumors consist of a heterogeneous population of cells with a subset population that has the exclusive tumorigenic ability. They are called cancer stem cells (CSCs). CSCs can self-renew to generate additional CSCs and also differentiate to generate phenotypically diverse cancer cells with limited proliferative potential. They have been identified in a variety of tumors. In this study, we identify the marker of CSCs in the established human laryngeal tumor Hep-2 cell line in vivo. Our in vitro experiment shown as CD133, a 5-transmembrane glycoprotein expressed in Hep-2 cell line. CD133 was supposed as a candidate of CSC in laryngeal carcinoma. In this study, the expression of CD133 was detected in a Hep-2 cell line. Applying the magnetic cell sorting (MACS) technology, we reported the results of purifying CD133 positive cells from a Hep-2 cell line. Three-type cells' tumor-forming ability was examined in vivo to identify the marker of CSCs in Hep-2 cell line.


METHODS
CD133 was selected as a putative marker of CSC in laryngeal carcinoma, Hep-2 cell lines. Flow cytometry was used to detect the expression of CD133 in the Hep-2 cell line. Immunomagnetic beads were applied to purify CD133-positive cells. CD133(+), CD133(-) tumor cells, and unsorted Hep-2 cells were injected into severe combined immune deficiency (SCID) mice individually to observe tumor-forming ability.


RESULTS
Only a small proportion (3.15% +/- 0.83%) of cells in the Hep-2 cell line express the CD133 marker. In comparison with CD133(-) tumor cells and unsorted cells, CD133(+) cells possess a marked capacity for tumor formation in vivo (p <.05).


CONCLUSIONS
CD133 is 1 of the markers for CSCs in human laryngeal tumors of the Hep-2 cell line. Work on the characterization of these cells provides a powerful tool to investigate the tumorigenic process in the larynx and to develop therapies targeting the CSC.
Department of Otolaryngology-Head and Neck Surgery, Fudan University, Affiliated Eye, Ear, Nose, and Throat Hospital, Shanghai, China.
Xu Dong
XD
Wei
Liang
L
Zhou
Lei
L
Cheng
Jie
J
Tian
Jack J
JJ
Jiang
Julia
J
Maccallum
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000302:isolation & purification / Q000378:metabolism*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D007822:Laryngeal Neoplasms / Q000378:metabolism*; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism*
D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse