Figure 7. IL-1α-stimulated PSCs increase HGF secretion and enhance cancer cell migration and DNA synthesis.

(A) HGF secretion was measured by ELISA in conditioned medium from pancreatic stellate cells SC40, SC41 and SC42, incubated in the presence or absence of IL-1α (1 ng/ml). (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 IL-1α (1 ng/ml). The cells were lysed and proteins subjected to immunoblotting using the indicated antibodies. Immunoblots shown are representative of three independent experiments. (C) The effect of IL-1α stimulated pancreatic stellate cells on cancer cell DNA synthesis was analysed by measuring [³H]-thymidine incorporation. BxPC-3 cells were stimulated for 24 hours with IL-1α (1 ng/ml) or with conditioned medium obtained from PSCs cultured in the presence or absence of IL-1α (1 ng/ml), with [³H]-thymidine added at 18 hours. The results are presented as mean +/−SEM normalized to unstimulated control. (D) BxPC-3 cells were cultured in colonies to confluence and scratch wounds were established in the centre of the colony. Control medium including IL-1α (1 ng/ml) or conditioned medium from PSC cultured in the presence or absence of IL-1α (1 ng/ml) was transferred to the wound assays. The wound area was measured at 0 and 10 h and normalized to controls. Direct effects of IL-1α were measured in independent experiments. Error bars represent S.E.M.; *p < 0.05, **p < 0.005, ***p < 0.001.