MRMT-1 rat breast carcinoma cells and models of bone metastases: improvement of an in vitro system to mimic the in vivo condition.

Breast cancer frequently metastasizes to the skeleton thus interrupting the normal bone remodeling process and causing bone degradation. Having suitable in vitro and in vivo models is important for understanding the pathogenesis and developing treatment strategies for bone metastasis in humans. In order to improve and characterize an in vitro model of bone metastasis from breast cancer an MRMT-1 rat breast carcinoma cell line or their conditioned medium were directly co-cultured with rat monocytes. To confirm the in vitro results, an in vivo model, in which MRMT-1 cells were inoculated into the proximal surface of the tibia, was also adopted. Osteoclast viability, activity and differentiation showed a significant increase (p<0.05, p<0.0005, p<0.0005, respectively) between co-culture with MRMT-1 cells and the other culture conditions. Moreover, scanning electron microscopy analysis, phalloidin staining and 4'-6-diamidino-2-phenylindole (DAPI) nuclear acid staining confirmed that co-culture with MRMT-1 cells also induced a greater differentiation in osteoclast structure and morphology. Finally, the in vivo outcome at 3 weeks showed the presence of a severe osteolytic lesion, thus confirming the effectiveness of the present in vitro results. These results demonstrated an improvement of an in vitro model of bone metastases from breast cancer in which co-culture with MRMT-1 cells was shown to be a dynamic system that closely mimics the in vivo situation. The present study may help improve our understanding of the complex "vicious cycle" between osteoblasts, osteoclasts and tumor cells.