Proteomic signature of reversine-treated murine fibroblasts by 2-D difference gel electrophoresis and MS : Possible associations with cell signalling networks

Recent advances in stem cell biology have demonstrated that terminally differentiated adult cells can be induced to de-differentiate into progenitor cells (induced stem cells) upon proper stimuli. This has been achieved by the induced expression of key regulatory genes by retro- or lenti-viral systems. On the other hand, synthetic "small molecules" can also induce de-differentiation and may represent a potentially safer approach as compared with genetic manipulation. Along this line, a synthetic purine called "reversine" has been shown to induce the de-differentiation of fibroblasts into mesenchymal stem-cell-like progenitors, which can be successively induced to differentiate into skeletal muscle, smooth muscle and bone cells. The mechanism whereby reversine is able to achieve de-differentiation has yet to be clarified. In this context, we defined the protein changes induced by reversine treatment in murine fibroblasts by 2-D difference gel electrophoresis, coupled with MS. Proteins involved in cytoskeletal and cell shape remodeling, RNA export, degradation, folding, stress control and ATP production were found to be remarkably changed after reversine treatment. Ingenuity pathway analysis (IPA) predicted that these protein pattern changes enabled to propose that about 40 proteins might be associated to several biological functional networks, including cellular assembly, cell signaling and cell death. Altogether our data confirm the intrinsic complexity of the de-differentiation process induced by reversine and suggest more selected approaches to investigate the action mechanism of this small molecule.