RESUMEN
Prostaglandin F2alpha (PGF2alpha) induces cyclin D1 expression and DNA synthesis in Swiss 3T3 cells. In order to assess which signaling mechanisms are implicated in these processes, we have used both a pharmacological approach and interfering mutants. We demonstrate that PGF2alpha induces extracellular-signal-regulated kinase (ERK1-2) and p38MAPK activation, and inhibition of any of these signaling pathways completely blocks PGF2alpha-stimulated DNA synthesis. We also show that ERK1-2, but not p38MAPK activation is required to induce cyclin D1 expression, strongly suggesting that the concerted action of cyclin D1 gene expression and other events are required to induce complete phosphorylation of retinoblastoma protein and S-phase entry in response to PGF2alpha.
Asunto(s)
División Celular/efectos de los fármacos , Dinoprost/farmacología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Animales , Western Blotting , Ciclina D1/genética , Replicación del ADN , Electroforesis en Gel de Poliacrilamida , Activación Enzimática , Ratones , Células 3T3 SwissRESUMEN
Leukaemia inhibitory factor (LIF) or Oncostatin M (OSM), both mitogens for Swiss mouse 3T3 cells, triggers initiation of DNA synthesis without the requirement for mevalonic acid. Thus, Lovastatin (LOV), an inhibitor of the hydroxy methylglutaryl CoA (HMGCoA) reductase, does not block LIF or OSM induced DNA replication and cell multiplication. In contrast, increasing concentrations of LOV from 1 to 60 microM block the mitogenic action of PGF(2alpha) by decreasing the number of cells capable of entering S-phase and dividing. This inhibition by LOV can be reversed by addition of mevanolactone (MEV), an analogue of mevalonic acid. Thus, LIF or OSM triggers initiation of DNA replication independently of mevalonic acid synthesis and therefore without the involvement of isoprenylation of various signalling proteins.