title: Identification of cancer stem-like side population cells in purified primary cultured human laryngeal squamous cell carcinoma epithelia.
abstract: Cancer stem-like side population (SP) cells have been identified in many solid tumors; however, most of these investigations are performed using established cancer cell lines. Cancer cells in tumor tissue containing fibroblasts and many other types of cells are much more complex than any cancer cell line. Although SP cells were identified in the laryngeal squamous cell carcinoma (LSCC) cell line Hep-2 in our pilot study, it is unknown whether the LSCC tissue contains SP cells. In this study, LSCC cells (LSCCs) were primary cultured and purified from a surgically resected LSCC specimen derived from a well-differentiated epiglottic neoplasm of a Chinese male. This was followed by the verification of epithelium-specific characteristics, such as ultrastructure and biomarkers. A distinct SP subpopulation (4.45±1.07%) was isolated by Hoechst 33342 efflux analysis from cultured LSCCs by using a flow cytometer. Cancer stem cell (CSC)-associated assays, including expression of self-renewal and CSC marker genes, proliferation, differentiation, spheroid formation, chemotherapy resistance, and tumorigenicity were then conducted between SP and non-SP (NSP) LSCCs. In vitro and in vivo assays revealed that SP cells manifested preferential expression of self-renewal and CSC marker genes, higher capacity for proliferation, differentiation, and spheroid formation; enhanced resistance to chemotherapy; and greater xenograft tumorigenicity in immunodeficient mice compared with NSP cells. These findings suggest that the primary cultured and purified LSCCs contain cancer stem-like SP cells, which may serve as a valuable model for CSC research in LSCC.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, People's Republic of China.
authors.forename: Chun-Ping
authors.initials: CP
authors.lastname: Wu
authors.forename: Liang
authors.initials: L
authors.lastname: Zhou
authors.forename: Ming
authors.initials: M
authors.lastname: Xie
authors.forename: Huai-Dong
authors.initials: HD
authors.lastname: Du
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
authors.forename: Shan
authors.initials: S
authors.lastname: Sun
authors.forename: Jin-Yan
authors.initials: JY
authors.lastname: Li
mesh_terms: D000368:Aged; D000818:Animals; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000473:pathology* / Q000648:ultrastructure; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D005434:Flow Cytometry; D006258:Head and Neck Neoplasms / Q000473:pathology* / Q000648:ultrastructure; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D046529:Microscopy, Electron, Transmission; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D058985:Side-Population Cells / Q000166:cytology* / Q000378:metabolism; D000077195:Squamous Cell Carcinoma of Head and Neck; D014407:Tumor Cells, Cultured