Prednisolone inhibits SaOS2 osteosarcoma cell proliferation by activating inducible nitric oxide synthase

• 作者: Alessandra Cazzaniga Jeanette AM Maier Sara Castiglioni
• 来源期刊: 世界转化医学杂志
• 学科: 医学
• 关键词: OSTEOSARCOMA cells PREDNISOLONE NITRIC OXIDE INDUCIBLE NITRIC OXIDE SYNTHASE Endothelial NITRIC OXIDE SYNTHASE Reactive oxygen species
• 年，卷（期）: sjzhyxzz_2016,（1）
• 页码范围: 53-58
• 页数: 6页
• 分类号: R

摘要

AIM:To investigate the effect of prednisolone,a synthetic glucocorticoid used in inflammatory diseases,on the growth of cultured osteosarcoma cells.METHODS:Two osteosarcoma cell lines with different degree of differentiation were used.SaOS2 show a rather mature phenotype,while U2OS are negative for almost all osteoblastic markers.The cells were exposed to different concentrations of prednisolone(1-9μmol/L)with or without antioxidants or the inhibitor of inducible nitric oxide synthase(iNOS)l-N6-(iminoethyl)-lysine-HCl(L-NIL).Cell growth was assessed by counting viable cells.The production of nitric oxide(NO)was measured in the conditioned media by the Griess method.The production of reactive oxygen species was quantified using 2’-7’-dichlorofluorescein diacetate.Western blot with specific antibodies against NOSs was performed on cell extracts.RESULTS:Prednisolone inhibited SaOS2 cell growth in a dose dependent manner.No significant effects were observed in U2OS.The inhibition of SaOS2 growth is not due to oxidative stress,because antioxidants do not rescue cell proliferation.Since high concentrations of NO inhibit bone formation,we also measured NO and found it induced in SaOS2,but not in U2OS,exposed to prednisolone,because of the upregulation of iNOS as detected by western blot.Therefore,we treated SaOS2 with prednisolone in the presence or in the absence of L-NIL.L-NIL prevented NO release induced by prednisolone at all the concentrations apart from 9μmol/L.At the same concentrations,we found that L-NIL rescued SaOS2 growth after exposure to prednisolone.In U2OS cells,prednisolone did not induce NO production nor affected cell growth.All together,these data indicate that a link exists between increased amounts of NO and growth inhibition in response to prednisolone in SaOS2.CONCLUSION:Prednisolone inhibited SaOS2 proliferation by increasing the release of NO through the upregulation of iNOS,while no effect was exerted on U2OS.