Cellular mechanism of metformin action

Activation of the insulin receptor tyrosine kinase and tyrosine phosphorylation of intracellular substrates are important steps in insulin signalling. In order to elucidate the cellular mechanism of action of metformin (NN'dimethylbiguanide) we have focused towards the effects of metformin on the insulin receptor kinase, the phosphorylation cascade and the biological effect of insulin. Since annexins (lipocortins) have been recently recognized as substrates of several tyrosine kinases we have investigated the effect of metformin on phosphorylation of annexins after insulin stimulation or microinjection of pp60c-src kinase in Xenopus laevis oocytes. Insulin induced in oocytes progression through the cell cycle from late G2 to M phase (maturation). Microinjection of pp60c-src kinase or treatment with metformin potentiates both the rate and the level of insulin-induced oocyte maturation. In oocytes prelabeled with 32P orthophosphate metformin potentiates insulin induced phosphorylation of annexins. It is concluded that annexins are substrates of the phosphorylation cascade initiated by insulin which is synergistic to the action of pp60c-src kinase and that this early phosphorylation events correlate well with the enhanced biological effect of insulin during metformin treatment.