Cirri P, Taddei M L, Chiarugi P, Buricchi F, Caselli A, Paoli P, Giannoni E, Camici G, Manao G, Raugei G, Ramponi G
Dipartimento di Scienze Biochimiche, Università di Firenze, 50134 Firenze, Italy.
Mol Biol Cell. 2005 Jan;16(1):73-83. doi: 10.1091/mbc.e04-01-0011. Epub 2004 Nov 3.
Cellular behavior can be considered to be the result of a very complex spatial and temporal integration of intracellular and extracellular signals. These signals arise from serum-soluble factors as well as from cell-substrate or cell-cell interactions. The current approach in mitogenesis studies is generally to analyze the effect of a single growth factor on serum-starved cells. In this context, a metabolic hormone such as insulin is found to be a mitogenic agent in many cellular types. In the present study, we have considered the effect of insulin stimulation in platelet-derived growth factor (PDGF)-activated NIH-3T3 and C2C12 cells. Our results show that insulin is able to inhibit strongly both NIH-3T3 and C2C12 cell growth induced by PDGF, one of the most powerful mitotic agents for these cell types. This inhibitory effect of insulin is due primarily to a premature down-regulation of the PDGF receptor. Thus, when NIH-3T3 or C2C12 cells are stimulated with both PDGF and insulin, we observe a decrease in PDGF receptor phosphorylation with respect to cells treated with PDGF alone. In particular, we find that costimulation with insulin leads to a reduced production of H2O2 with respect to cell stimulation with PDGF alone. The relative low concentration of H2O2 in PDGF/insulin-costimulated cell leads to a limited down-regulation of protein tyrosine phosphatases, and, consequently, to a reduced PDGF receptor phosphorylation efficiency. The latter is very likely to be responsible for the insulin-dependent inhibition of PDGF-receptor mitogenic signaling.
细胞行为可被视为细胞内和细胞外信号在时空上非常复杂整合的结果。这些信号源于血清可溶性因子以及细胞与底物或细胞与细胞之间的相互作用。目前有丝分裂研究的方法通常是分析单一生长因子对血清饥饿细胞的影响。在这种情况下,一种代谢激素如胰岛素在许多细胞类型中被发现是一种有丝分裂原。在本研究中,我们考虑了胰岛素刺激对血小板衍生生长因子(PDGF)激活的NIH-3T3和C2C12细胞的影响。我们的结果表明,胰岛素能够强烈抑制PDGF诱导的NIH-3T3和C2C12细胞生长,而PDGF是这些细胞类型中最强大的有丝分裂剂之一。胰岛素的这种抑制作用主要是由于PDGF受体的过早下调。因此,当用PDGF和胰岛素同时刺激NIH-3T3或C2C12细胞时,相对于仅用PDGF处理的细胞,我们观察到PDGF受体磷酸化水平降低。特别是,我们发现与单独用PDGF刺激细胞相比,胰岛素共刺激导致H2O2的产生减少。在PDGF/胰岛素共刺激的细胞中相对较低浓度的H2O2导致蛋白酪氨酸磷酸酶的下调受限,因此导致PDGF受体磷酸化效率降低。后者很可能是胰岛素依赖性抑制PDGF受体有丝分裂信号传导的原因。