title: CD133 expression and identification of CD133/nestin positive cells in rhabdomyosarcomas and rhabdomyosarcoma cell lines.
abstract: BACKGROUND
  Co-expression of CD133, cell surface glycoprotein, and nestin, an intermediate filament protein, was determined to be a marker of neural stem cells and of cancer stem cells in neurogenic tumors.
  
  
  METHODS
  We examined the expression of CD133 and nestin in ten tumor tissue samples taken from patients with rhabdomyosarcomas and in five rhabdomyosarcoma cell lines. Immunohistochemistry and immunofluorescence were used to examine FFPE tumor tissue samples. Cell lines were analyzed by immunofluorescence, immunoblotting, flow cytometry, and RT-PCR. Functional assays (clonogenic in vitro assay and tumorigenic in vivo assay) were also performed using these cell lines.
  
  
  RESULTS
  CD133 and nestin were detected in all 10 tumor tissue samples and in all 5 cell lines; however, the frequency of CD133+, Nes+, and CD133+/Nes+ cells, as well as the intensity of fluorescence varied in individual samples or cell lines. The expression of CD133 and nestin was subsequently confirmed in all cell lines by immunoblotting. Furthermore, we observed an increasing expression of CD133 in relation to the cultivation. All cell lines were positive for Oct3/4 and nucleostemin; NSTS-11 cells were also able to form xenograft tumors in mice.
  
  
  CONCLUSIONS
  Our results represent the first evidence of CD133 expression in rhabdomyosarcoma tissue and in rhabdomyosarcoma cell lines. In addition, the co-expression of CD133 and nestin as well as results of the functional assays suggest a possible presence of cancer cells with a stem-like phenotype in these tumors.
authors.affiliation: Laboratory of Tumor Biology and Genetics, Department of Experimental Biology, School of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Sana
authors.forename: Iva
authors.initials: I
authors.lastname: Zambo
authors.forename: Jan
authors.initials: J
authors.lastname: Skoda
authors.forename: Jakub
authors.initials: J
authors.lastname: Neradil
authors.forename: Petr
authors.initials: P
authors.lastname: Chlapek
authors.forename: Marketa
authors.initials: M
authors.lastname: Hermanova
authors.forename: Peter
authors.initials: P
authors.lastname: Mudry
authors.forename: Alzbeta
authors.initials: A
authors.lastname: Vasikova
authors.forename: Karel
authors.initials: K
authors.lastname: Zitterbart
authors.forename: Ales
authors.initials: A
authors.lastname: Hampl
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Sterba
authors.forename: Renata
authors.initials: R
authors.lastname: Veselska
mesh_terms: D000071916:AC133 Antigen; D000293:Adolescent; D015703:Antigens, CD / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D005260:Female; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D064231:Nestin; D010455:Peptides / Q000378:metabolism*; D012208:Rhabdomyosarcoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D013234:Stem Cells / Q000378:metabolism; D055815:Young Adult
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D014408:Biomarkers, Tumor; D006023:Glycoproteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse