Targeted Knockdown of Tissue Factor in B16F10 Melanoma Cells Suppresses their Ability to Metastasize to Bone and Cause Cancellous Bone Loss

Amanda Politano, Martin Butcher, Melec Zeadin, Peter Gross, Nima Vaezzadeh, and Stephen G. ShaughnessyDepartment of Pathology and Molecular Medicine, McMaster University and the Thrombosis & Atherosclerosis Research Institute, Hamilton, Ontario, Canada. AbstractIn this study, we use a well-defined mouse model to examine tissue factor's (TF) role in osteolytic bone metastasis. C57BL/6 mice received either mock siRNA-transfected or TF-specific siRNA-transfected B16F10 melanoma cells by left ventricular injection. A third group served as an age-matched control and did not receive any tumour cells. The effect on tumour burden and bone strength was then determined 14 days later by using bone histomorphometry and biomechanical testing. Based on histomorphometric analysis of the femurs, mice receiving TF-specific siRNA-transfected tumour cells had significantly reduced tumour burden as compared to those from mice that received mock siRNA-transfected tumour cells (2.20 ± 0.58% vs. 9.18 ± 2.20%). Furthermore, the femurs from mice receiving TF siRNA-transfected tumour cells displayed decreased osteoclast surface and consequently, increased cancellous bone volume and strength when compared to those isolated from mice that were injected with mock-transfected tumour cells. More importantly, no differences in osteoclast surface or cancellous bone volume and strength were observed when the femurs of mice that received TF siRNA-transfected tumour cells were compared to control mice that did not receive tumour cells. Based on these findings, we conclude that the expression of TF by tumour cells promotes their ability to metastasize to bone, thereby facilitating tumour cell-induced cancellous bone loss.