Figure 5. c-Met signaling promotes perivascular growth and angiogenesis of cancer cells.
(A) 231BrM/Dox/sh-c-Met and SKBrM3/Dox/sh-c-Met cells were treated with or without doxycycline to induce knockdown of c-Met for 48hrs and the cells were seeded on top of HBMECs. Cells were washed with PBS and the number of attached GFP+ tumor cells were counted under a fluorescent microscope. (B) HBMECs (2 × 104) were labeled with red cell tracker dye and seeded on top of the matrigel for 6 hrs to form the tube-like structure. 5 × 103 tumor cells labeled with the green cell tracker dye were treated with or without doxycycline for 48hrs and they were seeded on top of the HBMECs. Photos were taken after 6hrs incubation. Scale bars, 20 µm. (C) 2 × 104 HBMECs were incubated in the indicated conditioned medium with or without 10mM repertaxin for 6hrs followed by measuring their tube forming abilities. (D) 2 × 104 HBMECs were incubated in the conditioned medium prepared from 231BrM/Dox/sh-c-Met and SKBrM3/Dox/sh-c-Met cells with or without the treatment of doxycycline. Photos were taken after 6hrs of incubation followed by measuring the tube forming ability of HBMECs. (E) 2 × 104 HBMECs were incubated with the conditioned medium prepared from MCF7 or MDA453 with or without ectopic expression of c-Met. Photos were taken after 6hrs incubation followed by measuring the tube forming ability of HBMECs. Scale bars, 100 µm.