Figure 8. TGFβ-stimulated PSCs reduces HGF secretion and enhance cancer cell migration and DNA synthesis.

(A) HGF secretion was measured by ELISA in conditioned medium from the pancreatic stellate cells SC40, incubated in the presence or absence of TGFβ (2 ng/ml). The results are normalized to unstimulated controls. (B) BxPC-3 cells were stimulated for the indicated time points with conditioned medium from the PSCs (SC40) cultured in the presence or absence of TGFβ (2 ng/ml). The cells were lysed and proteins subjected to immunoblotting using the indicated antibodies. Immunoblots shown are representative of three independent experiments. (C) DNA synthesis was determined by measuring [³H]-thymidine incorporation in BxPC-3 cells stimulated with TGFβ (2 ng/ml) or conditioned medium from the pancreatic stellate cells SC40, incubated in the presence or absence of TGFβ (2 ng/ml). TGFβ and PSC conditioned medium were added at 48 h, [³H]-thymidine after 68 h and the cells were harvested at 72 h. (D) Cancer cell migration was determined by wound assay after 10 h stimulation of BxPC-3 cells with TGFβ or conditioned medium from the pancreatic stellate cells SC42, SC41 and SC40, incubated in the presence or absence of TGFβ (2 ng/ml). The wound area was measured at 0 and 10 h and normalized to controls. Error bars represent S.E.M.; *p < 0.05, **p < 0.005, ***p < 0.001.