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AIM:To investigate the effects of transforming growth factorβ1(TGF-β1)on the differentiation of colonic lamina propria fibroblasts(CLPF)into myofibroblasts in vitro. METHODS:Primary CLPF cultures were incubated with TGF-β1 and analyzed for production ofα-smooth muscle actin(α-SMA),fibronectin(FN)and FN isoforms.Migration assays were performed in a modified 48-well Boyden chamber.Levels of total and phosphorylated focal adhesion kinase(FAK)in CLPF were analyzed after induction of migration.RESULTS:Incubation of CLPF with TGF-β1 for 2 d did not changeα-SMA levels,while TGF-β1 treatment for 6 d significantly increasedα-SMA production. Short term incubation(6 h)with TGF-β1 enhanced CLPF migration,while long term treatment(6 d)of CLPF with TGF-β1 reduced migration to 15%-37% compared to untreated cells.FN and FN isoform mRNA expression were increased after short term incubation with TGF-β1(2 d)in contrast to long term incubation with TGF-β1 for 6 d.After induction of migration, TGF-β1-preincubated CLPF showed higher amounts of FN and its isoforms and lower levels of total and phosphorylated FAK than untreated cells. CONCLUSION:Long term incubation of CLPF with TGF-β1 induced differentiation into myofibroblasts with enhancedα-SMA,reduced migratory potential and FAK phosphorylation,and increased FN production.In contrast,short term contact(6 h)of fibroblasts with TGF-β1 induced a dose-dependent increase of cell migration and FAK phosphorylation without induction ofα-SMA production.
METHODS: Primary investigate the effects of transforming growth factor β1 (TGF-β1) on the differentiation of colonic lamina propria fibroblasts (CLPF) into myofibroblasts in vitro. Methods: Primary CLPF cultures were incubated with TGF-β1 and analyzed for production of α-smooth muscle Leaks of total and phosphorylated focal adhesion kinase (FAK) in CLPF were analyzed after induction of migration .RESULTS: (a-SMA), fibronectin (FN) and FN isoforms. Migration assays were performed in a modified 48-well Boyden chamber. Incubation of CLPF with TGF-β1 for 2 d did not alter α-SMA levels while TGF-β1 treatment for 6 d significantly increased α-SMA production. Short term incubation (6 h) with TGF-β1 enhanced CLPF migration, while long term treatment (6 d) of CLPF with TGF-β1 reduced migration to 15% -37% compared to untreated cells. NF and FN isoform mRNA expression were increased after short term incubation with TGF-β1 (2 d) in contrast to long term incubation with TGF-β1 for 6 d. After induction of migration, TGF-β1-preincubated CLPF showed higher amounts of FN and its isoforms and lower levels of total and phosphorylated FAK than untreated cells. CONCLUSION: Long term incubation of CLPF with TGF-β1 induced differentiation into myofibroblasts with enhanced α-SMA, reduced migratory potential and FAK phosphorylation, and increased FN production. Contrast, short term contact (6 h) of fibroblasts with TGF-β1 induced a dose-dependent increase of cell migration and FAK phosphorylation without induction of α-SMA production.