Cellular and molecular phenotypes of proliferating stromal cells from human carcinomas.
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.
Kopantzev
E P
EP
M M Shemyakin and Yu A Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia. kopantzev@ibch.ru
Vayshlya
N A
NA
Kopantseva
M R
MR
Egorov
V I
VI
Pikunov
M
M
Zinovyeva
M V
MV
Vinogradova
T V
TV
Zborovskaya
I B
IB
Sverdlov
E D
ED
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
C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins
Hum Mutat. 2003 Mar;21(3):229-39
12619108
Exp Cell Res. 2001 Mar 10;264(1):169-84
11237532
Nat Rev Cancer. 2004 Nov;4(11):839-49
15516957
Clin Cancer Res. 1998 Jun;4(6):1527-32
9626473
Am J Clin Oncol. 2004 Aug;27(4):425-8
15289739
Exp Mol Pathol. 1990 Apr;52(2):221-34
2185029
Nat Rev Cancer. 2008 Jan;8(1):61-70
18075512
Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13180-5
16924102
Curr Opin Genet Dev. 2001 Feb;11(1):54-9
11163151
Cancer Res. 2006 Apr 1;66(7):3392-5
16585159
J Cell Sci. 2005 Feb 1;118(Pt 3):485-96
15657080
Cell. 2005 Dec 16;123(6):1001-11
16360031
Cell. 2008 May 16;133(4):704-15
18485877
J Clin Invest. 1995 Feb;95(2):859-73
7532191
Chest. 1994 Dec;106(6 Suppl):377S-381S
7988268
Nat Rev Cancer. 2006 May;6(5):392-401
16572188
Nature. 2004 Nov 18;432(7015):332-7
15549095
Clin Cancer Res. 1998 Dec;4(12):2931-8
9865903
J Clin Oncol. 1999 Jul;17(7):2100-4
10561264
Ann N Y Acad Sci. 2004 Apr;1014:155-63
15153430
J Cell Biol. 1996 Jul;134(1):67-80
8698823
Clin Cancer Res. 1999 May;5(5):991-1000
10353731
Nat Rev Mol Cell Biol. 2003 Aug;4(8):657-65
12923528
Am J Respir Cell Mol Biol. 2008 Jan;38(1):95-104
17673689
Nat Rev Cancer. 2002 Jun;2(6):442-54
12189386
Clin Exp Metastasis. 2008;25(6):621-8
18600305
Anal Biochem. 1990 Nov 15;191(1):31-4
1706565
J Clin Invest. 2002 Aug;110(3):341-50
12163453
Cancer Res. 2005 Mar 1;65(5):1627-30
15753354