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Fig. 7. The C-terminus of ephrin-B1 regulates the invasion of cancer cells. (A) Wild-type (WT) ephrin-B1 or C4 ephrin-B1 mutant was expressed in Capan-1 cells. The cells were seeded onto a Transwell membrane coated with a collagen matrix (25 µg/cm²) in serum-free medium containing control Fc or EphB2-Fc (4 µg/ml) with or without addition of the MMP-8 inhibitor (1 µM). In the lower chamber, medium containing 5% FBS was added as a chemoattractant. After 8 hours incubation, the wells were harvested and cells that had invaded the collagen were counted. Representative fields are shown. (Right) The results from three independent experiments, each in duplicate, are shown as the mean ± s.d. ^*P<0.01. (B) PANC-1 ephrin-B1 cells or PANC-1 cells transfected with a mock vector (mock) were injected intraperitoneally into nude mice. The representative appearance of intestinal loops 8 weeks after injection is shown. Arrows indicate disseminated tumor nodules in the mesentery. The right panel shows the histology of the tumors in the mesentery (x100). The asterisk indicates a tumor nodule. Microscopic invasion of cancer cells was observed in the mesenteric sheet (blanket). (C) Representative appearance of the tumors of panc1 cells expressing either mock vector, wild-type or C4 ephrin-B1 in the rectouterine region was compared. Yellow and red arrowheads indicate uterine horns and tumor nodules, respectively.

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