Inhibitors of Calcineurin Block Expression of Cyclins A and E Induced by Fibroblast Growth Factor in Swiss 3T3 Fibroblasts

In Swiss 3T3 fibroblasts, growth factor-stimulated progression from G1 to S phase involves activation of the Ca2+/calmodulin-dependent serine/threonine-specific protein phosphatase 2B (calcineurin). Here we report that both cobalt and the calcium chelator EGTA, inhibitors of calcium uptake, as well as cyclosporin A and FK-506, specific inhibitors of calcineurin function, abolished fibroblast growth factor (FGF)-induced expression of cyclins A and E, but not cyclin D1. At 0.1 μM concentration cyclosporin A completely blocked FGF-induced expression of cyclins E and A and it inhibited FGF-stimulated DNA synthesis by 40%; full inhibition of DNA synthesis required 10 μM cyclosporin A. PD 98059, an inhibitor of mitogen-activated protein (MAP) kinase kinase, and hemicholinium-3, an inhibitor of FGF-induced MAP kinase activity, did not inhibit the stimulatory effect of FGF on the expression of cyclin E. On the other hand, the inhibitory effect of 0.1 μM cyclosporin A on FGF-stimulated DNA synthesis was additive with that of hemicholinium-3, suggesting that the two inhibitors acted by different mechanisms. The inhibitors of calcineurin and calcium uptake also completely blocked the stimulatory effects of lysophosphatidic acid on the expression of cyclins E and A, but not cyclin D1. The results suggest that FGF- or lysophosphatidic acid-induced transcription of cyclin A and cyclin E genes is mediated by calcineurin involving a MAP kinase-independent mechanism and that increased expression of cyclins A and E is required for the maximal stimulatory effects of these mitogens on DNA synthesis.