title: Characterization of Cancer Stem Cells in Moderately Differentiated Buccal Mucosal Squamous Cell Carcinoma.
abstract: OBJECTIVE
  To identify and characterize cancer stem cells (CSC) in moderately differentiated buccal mucosa squamous cell carcinoma (MDBMSCC).
  
  
  METHODS
  Four micrometer-thick, formalin-fixed, paraffin-embedded MDBMSCC samples from six patients underwent 3,3-diaminobenzidine (DAB) immunohistochemical (IHC) staining for the embryonic stem cell (ESC) markers, NANOG, OCT4, SALL4, SOX2, and pSTAT3; cancer stem cell marker, CD44; squamous cell carcinoma (SCC) marker, EMA; and endothelial marker, CD34. The transcriptional activities of the genes encoding NANOG, OCT4, SOX2, SALL4, STAT3, and CD44 were studied using NanoString gene expression analysis and colorimetric in situ hybridization (CISH) for NANOG, OCT4, SOX2, SALL4, and STAT3.
  
  
  RESULTS
  Diaminobenzidine and immunofluorescent (IF) IHC staining demonstrated the presence of (1) an EMA(+)/CD44(+)/SOX2(+)/SALL4(+)/OCT4(+)/pSTAT3(+)/NANOG(+) CSC subpopulation within the tumor nests; (2) an EMA(-)/CD44(-)/CD34(-)/SOX2(+)/OCT4(+)/pSTAT3(+)/NANOG(+) subpopulation within the stroma between the tumor nests; and (3) an EMA(-)/CD44(-)/CD34(+)/SOX2(+)/SALL4(+)/OCT4(+)/pSTAT3(+)/NANOG(+) subpopulation on the endothelium of the microvessels within the stroma. The expression of CD44, SOX2, SALL4, OCT4, pSTAT3, and NANOG was confirmed by the presence of mRNA transcripts, using NanoString analysis and NANOG, OCT4, SOX2, SALL4, and STAT3 by CISH staining.
  
  
  CONCLUSIONS
  This study demonstrated a novel finding of three separate CSC subpopulations within MDBMSCC: (1) within the tumor nests expressing EMA, CD44, SOX2, SALL4, OCT4, pSTAT3, and NANOG; (2) within the stroma expressing SOX2, SALL4, OCT4, pSTAT3, and NANOG; and (3) on the endothelium of the microvessels within the stroma expressing CD34, SOX2, SALL4, OCT4, pSTAT3, and NANOG.
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Helen H
authors.initials: HH
authors.lastname: Yu
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Therese
authors.initials: T
authors.lastname: Featherston
authors.affiliation: Gillies McIndoe Research Institute, Wellington, New Zealand; Wellington Regional Plastic, Maxillofacial and Burns Unit, Hutt Hospital, Wellington, New Zealand.
authors.forename: Swee T
authors.initials: ST
authors.lastname: Tan
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Alice M
authors.initials: AM
authors.lastname: Chibnall
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Helen D
authors.initials: HD
authors.lastname: Brasch
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Paul F
authors.initials: PF
authors.lastname: Davis
authors.affiliation: Gillies McIndoe Research Institute , Wellington , New Zealand.
authors.forename: Tinte
authors.initials: T
authors.lastname: Itinteang
keywords: buccal; cancer; mucosal; oral cavity; squamous cell carcinoma; stem cells