Figure 2.

The CaMKII antagonist, KN-93, arrests the growth of human osteosarcoma cells. (a) MG-63 cells were treated with either DMSO (control), KN-92 (10 μM) or KN-93 (10 μM) for 24 h. Cells were then lysed and protein extracts were separated by SDS-PAGE. Western blots were conducted using antibodies directed against p-α-CaMKII or actin. (b) MG-63 or 143B cells were cultured for 24 h and then treated for 24 and 48 h with DMSO (0.1% v/v) (control), KN-92 (10 μM) or KN-93 (10 μM). At the end of the study, an MTT proliferation assay was performed. Values were obtained from three separate experiments and represent the mean±s.e. *P<0.01. (c) MG-63 Cells were stained with PI and flow cytometric analysis of the DNA content was performed. Analysis of the cell cycle was performed using ModFit LT 3.1 software. Values were obtained from three separate experiments and represent the mean±s.e. *P<0.01. (d) MG-63 cells were seeded in growth factor-reduced Matrigel, supplemented with DMSO, 10 μM KN-92 or 10μM KN-93. Medium and treatment were replaced every day for 7 days. Arrows indicate tumor colonies. Photographs are representative of three different experiments. Colonies were counted from three randomly selected microscopic fields from three different experiments.