title: Cancer stem cell, cytokeratins and epithelial to mesenchymal transition markers expression in oral squamous cell carcinoma derived from ortothopic xenoimplantation of CD44highcells.
abstract: Oral squamous cell carcinoma (OSCC) is the most prevalent neoplasia of oral cavity worldwide and prognosis remains unchanged in decades. Recently, different authors reported that head and neck squamous cell carcinomas have a subpopulation of tumor initiating cells that apparently correspond to cancer stem cells (CSC) and are also responsible for tumor growth and metastasis. The purpose of the present study was to investigate the microscopic and phenotypic characteristics of OSCC tumors induced after orthotopic xenoimplantation of SCC9WTcell line and CSC-enriched subpopulation isolated from SCC9 cell line based on high expression of the putative CSC marker CD44. Different numbers of FACS-sorted SCC9 CD44highand CD44lowcells as well as SCC9WT(wild type) were transplanted into the tongue of BALB/C nude (NOD/SCID) mice to evaluate their tumorigenic potential. Sixty days post-induction, tumors were morphologically characterized and immunostained for CSC markers (CD44, Nanog and Bmi-1), epithelial-mesenchymal transition (Snail, Slug) and epithelial differentiating cell markers (cytokeratins 4, 13, 15, 17 and 19), as well as E-cadherin and β-catenin. The data presented here shows that SCC9 CD44highcells have higher ability to form tumors than SCC9 CD44lowcells, even when significantly lower numbers of SCC9 CD44highcells were transplanted. Immunoassessment of tumors derived from SCC9 CD44highcells revealed high expression of cytokeratin CK19, β-catenin, E-cadherin and CD44, and negative or low expression of CK17, CK4, CK15, CK13, Nanog, Bmi-1, Snail and Slug. While tumors derived from SCC9WTshowed high expression of CK17, CK19, CD44, Nanog, Bmi-1, Snail and Slug, and negative or low expression of CK4, CK15, CK13, β-catenin and E-cadherin. Thus, SCC9 CD44highcells were highly tumorigenic, capable of originating heterogeneous tumors and these tumors have a immunohistochemical profile different from those formed by the wild type cell line.
authors.affiliation: Department of Oral Pathology, School of Dentistry, University of São Paulo, São Paulo, Brazil.
authors.forename: Nathália Paiva
authors.initials: NP
authors.lastname: de Andrade
authors.affiliation: Department of Oral Pathology, School of Dentistry, University of São Paulo, São Paulo, Brazil.
authors.forename: Maria Fernanda Setúbal Destro
authors.initials: MF
authors.lastname: Rodrigues
authors.affiliation: Department of Biological Sciences, School of Dentistry, University of São Paulo, Bauru, Brazil.
authors.forename: Camila Oliveira
authors.initials: CO
authors.lastname: Rodini
authors.affiliation: Department of Oral Pathology, School of Dentistry, University of São Paulo, São Paulo, Brazil. Electronic address: fadnunes@usp.br.
authors.forename: Fabio Daumas
authors.initials: FD
authors.lastname: Nunes
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D006801:Humans; D007633:Keratins / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009062:Mouth Neoplasms / Q000378:metabolism* / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D000071250:Snail Family Transcription Factors / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C000604429:Snai1 protein, mouse; C000604449:Snai2 protein, mouse; D000071250:Snail Family Transcription Factors; D007633:Keratins
keywords: CD44 expression; Cancer stem cells; Fluorescence-activated cell sorting; Oral squamous cell carcinoma; Xenoimplantation