Figure 3.

Effects of daily systemic administration of 100 μg/kg rhBMP7 on the growth of human prostate cancer cells in bone marrow (a–d) and bone metastasis (e and f) in vivo after intraosseous transplantation or intracardiac inoculation of human PC-3M-Pro4/luc⁺ cells using whole body bioluminescent reporter imaging. a: BMP7 treatment of PC-3M-Pro4/luc⁺ cells growing in bone marrow of nude mice as monitored by BLI. b: Representative examples of bioluminescent photon emission in a vehicle- and BMP7-treated animal at day 21. c: Representative radiographs at day 21 after intraosseous transplantation of PC-3M-Pro4/luc⁺ cells growing in bone marrow (arrows indicate bone lesions in tibiae of nude mice). d: Immunohistochemical analyses of phosho-Smad1 localization at day 21 after intraosseous transplantation of PC-3M-Pro4/luc⁺ cells growing in bone marrow. The animals were either vehicle-treated (upper left panel) or treated with rhBMP7 (top right) 6 hours before explantation and fixation of the tibiae. The number of phospho-Smad1-positive cells in the BMP7-treated group is increased significantly. e: Effect of BMP7 treatment on the formation and growth of bone metastases by PC-3M-Pro4/luc⁺ in nude mice as monitored by BLI (tumor burden, number of bone metastases, representative examples of BLI at day 19). Relative light units (±SEM) emitted was quantified at least once weekly starting at day 0 after intraosseous implantation or intracardiac inoculation of the prostate cancer cells. n = 8 for each experimental group for intraosseous transplantation experiment (a–d); n = 10 for intracardiac inoculation (e). Values are expressed as means ± SEM. *P < 0.05, **P < 0.001 (analysis of variance).