title: Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas.
abstract: BACKGROUND
  Stromal cells are a functionally important component of human carcinomas. The aim of this study was to obtain and characterise primary cultures of stromal cells from human carcinomas and the corresponding surrounding normal tissue.
  
  
  METHODS
  Primary stromal cell cultures from tumours of lung, oesophagus and pancreas were obtained using a mild tissue dissociation method and a medium for culturing mesenchymal cells. Immunofluorescence staining and western blotting were used to analyse the expression of differentiation markers and selected known oncoproteins in the cell cultures obtained.
  
  
  RESULTS
  A panel of stromal primary cultures was prepared from different human tumours and from matched normal cancer-free tissues. The in vitro proliferative potential of tumour-associated fibroblasts was shown to be higher than that of matched normal stromal cells. A mutational analysis of the TP53 and KRAS2 genes in a number of stromal cultures did not reveal known mutations in most cells of the cultures studied. Western blot analysis showed that stromal cells of lung tumours were characterised by a statistically significantly lower expression level of the p16 protein as compared with that in normal lung stromal cells. An important finding of our study was that, according to immunofluorescence assay, a fraction of fibroblast-like vimentin-positive cells in some tumour and normal stromal cell cultures expressed an epithelial marker - cytokeratins.
  
  
  CONCLUSIONS
  Proliferating stromal cells from the carcinomas studied proved to be genetically normal cells with altered expression profiles of some genes involved in carcinogenesis, as compared with normal stromal cells. Epithelial-mesenchymal transition may lead to the emergence of transdifferentiated fibroblast-like cells in tumour stroma and in the tumour-surrounding tissue.
authors.affiliation: M M Shemyakin and Yu A Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia. kopantzev@ibch.ru
authors.forename: E P
authors.initials: EP
authors.lastname: Kopantzev
authors.forename: N A
authors.initials: NA
authors.lastname: Vayshlya
authors.forename: M R
authors.initials: MR
authors.lastname: Kopantseva
authors.forename: V I
authors.initials: VI
authors.lastname: Egorov
authors.forename: M
authors.initials: M
authors.lastname: Pikunov
authors.forename: M V
authors.initials: MV
authors.lastname: Zinovyeva
authors.forename: T V
authors.initials: TV
authors.lastname: Vinogradova
authors.forename: I B
authors.initials: IB
authors.lastname: Zborovskaya
authors.forename: E D
authors.initials: ED
authors.lastname: Sverdlov
mesh_terms: D000328:Adult; D000368:Aged; D015153:Blotting, Western; D002277:Carcinoma / Q000235:genetics* / Q000473:pathology*; D002460:Cell Line; D004252:DNA Mutational Analysis; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D020869:Gene Expression Profiling; D016158:Genes, p53; D006801:Humans; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D017154:Stromal Cells / Q000166:cytology*; D018631:ras Proteins / Q000235:genetics
chemical_list: C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins
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