Inhibition of protein kinase C activity inhibits osteosarcoma metastasis

Introduction: For some cancers bone is the preferred site for metastasis and involves a cascade involving transition of epithelial cells to mesenchymal cells and subsequent intravasation to the blood and lymph vessels, and finally hematogenous dissemination to perivascular niches of the bone marrow sinusoids. It has been shown that protein kinase C can aid metastasis to bone. Hence, pharmacological inhibition of protein kinase C (PKC) activity is thought of as a potential therapeutic option in bone metastatic lesions. The objective of the current study was to investigate how PKCs exert their effect on bone cancer metastasis and to test the efficacy of pharmacological inhibition of PKC on bone metastasis. Material and methods: The effect of the PKC inhibitor Go6983 on epithelial and mesenchymal cell marker expression in the osteosarcoma cell line DAN was determined by immunoblot and immunofluorescence analysis. The in vivo effect of Go6983 was evaluated with a xenograft model using DAN cells. Results: Treatment with transforming growth factor beta (TGF-beta) led to loss of the epithelial cell marker and gain of mesenchymal cell markers in the osteosarcoma cell line, DAN. This transition occurred concomitantly with PKC activation. TGF-beta-mediated PKC activation resulted in activation of ribosomal protein 6 (S6), but not S6K1. Pharmacological inhibition of PKC activation attenuated these effects. In a xenograft model of experimental metastasis, pharmacological inhibition of PKC activation over a period of 4 weeks reduced both tumor burden and metastasis to lungs. Conclusions: Our results indicate that PKC potentiates tumor metastasis to the bone by potentiating translation increase and can be putatively inhibited by pharmacological inhibition.