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Abstract
Scirrhous gastric cancer cells proliferate rapidly with fibrosis, when the cancer cells invade into the submucosa of the stomach. To investigate the mechanisms responsible for the rapid proliferation, the growth interaction between gastric cancer cells and fibroblasts was examined. Human gastric cancer cell lines established from scirrhous carcinoma or well-differentiated adenocarcinoma were used. Human fibroblast cell lines were obtained from various organs. The growth interaction between gastric cancer cells and fibroblasts was examined by calculating the number of cancer cells or by measuring [3H]thymidine incorporation of cancer cells. Gastric fibroblasts specifically stimulated the growth of scirrhous gastric cancer cells, but not that of well-differentiated adenocarcinoma cells. The growth factor(s) produced from gastric fibroblasts were then partially purified and characterised. The growth-promoting factor(s) had apparent molecular weights of 10000 dalton and was sensitive both to heat and proteinase treatment. No inhibition for the factor(s) was achieved with defined anti-growth factor antibodies. In this study, differential responses of scirrhous and well-differentiated gastric cancer cells to orthotopic fibroblasts were shown. Rapid proliferation of scirrhous gastric carcinoma should be partly controlled by orthotopic fibroblasts. The growth factor(s) from gastric fibroblasts, which was distinct from various defined growth factors such as epidermal growth factor (EGF), basic fibroblast growth factor (b-FGF), transforming growth factor-alpha (TGF-alpha), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), insulin-like growth factor I (IGF-I), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF) and transforming growth factor beta 1 (TGF-beta 1) may play an important role in the progression of scirrhous gastric cancer cells.
Highlights
Human fibroblast cell lines were obtained from various organs
We examined the effect of various defined growth factors, including epidermal growth factor (EGF) (Gibco), basic fibroblast growth factor (b-FGF) (Austral Biologics, San Ramon, CA, USA) keratinocyte growth factor (KGF) (UBI, Lake Placid, NY, USA), vascular endothelial growth factor (VEGF) (Pepro Tec, Rocky Hill, NJ, USA), transforming growth factor-a (TGF-a) (Becton Dickinson Labware, Mountain View, CA, USA), insulin-like growth factor I (IGF-I) (Mallinckrodt, St Louis, MO, USA), platelet-derived growth factor AA homodimer (PDGF-AA) (Austral Biologicals), hepatocyte growth factor (HGF) (Becton Dickinson Labware), and TGF,B1 (King Brewing, Kakogawa, Japan) which are thought to affect the growth of gastric carcinoma (Yoshida et al, 1989, 1990; Yasui et al, 1988; Tanimoto et al, 1991; Hattori et al, 1994; Shibamoto et al, 1992; Ito et al, 1992) on the growth of OCUM-2M cells
Growth-promoting activity of serum-free conditioned medium from fibroblasts for various gastric cancer cells The activity for OCUM-2M cells was evident following the addition of 25% Serum-free conditioned medium (SF-CM) with 1-2% fetal calf serum (FCS) (Figure 1); the
Summary
The growth-promoting activity of the treated SF-CM was determined by calculating the number of OCUM-2M cells. Aliquots of 250,l of each fraction were added to 750 pl of OCUM-2M cell suspension (1 x 104 cells per well) with 2% FCS in each well of 24-well dishes, and growth-promoting activity was determined by calculating the number of cancer cells. Effect of anti-growth factor antibodies on the growth activity of conditioned medium from fibroblasts
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