Kadri Z, Petitfrère E, Boudot C, Freyssinier J M, Fichelson S, Mayeux P, Emonard H, Hornebeck W, Haye B, Billat C
Laboratoire de Biochimie, Centre National de la Recherche Scientifique Formation de Recherche en Evolution, Institut Fédératif de Recherche 53 Biomolécules, UFR Sciences Exactes et Naturelles, UFR de Medecine, Université de Re.
Cell Growth Differ. 2000 Nov;11(11):573-80.
In the present study, we demonstrate that erythropoietin (Epo) induces the expression and the release of tissue inhibitors of metalloproteinase-1 (TIMP-1) in a time- and dose-dependent manner in Epo-dependent cell line UT-7 cells and in normal human erythroid progenitor cells from cord blood (CD36+) and required de novo protein synthesis. TIMP-1 was not expressed in the absence of Epo. Inhibition of the mitogen-activated protein kinase pathway by the specific inhibitors PD98059 and U0126 and of phosphatidylinositol 3-kinase by LY294002, strongly inhibited Epo-induced TIMP-1 expression and secretion. In the absence of Epo, both latent and active forms of matrix metalloproteinase-9 (MMP-9) were secreted into media. Upon Epo stimulation, MMP-9 and pro-MMP-9 secretion was inhibited in a dose-dependent manner parallel to TIMP-1 induction. The addition of PD98059, U0126, and LY294002 in the presence of Epo restored MMP-9 production in UT-7 and CD36+ cells. Our findings strongly suggest an inversely coordinated regulation of the TIMP-1 gene and MMP-9 production by Epo via mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways.
在本研究中，我们证明促红细胞生成素（Epo）在依赖Epo的细胞系UT-7细胞以及来自脐血的正常人红系祖细胞（CD36+）中，以时间和剂量依赖的方式诱导金属蛋白酶组织抑制剂-1（TIMP-1）的表达和释放，且这一过程需要从头合成蛋白质。在没有Epo的情况下，TIMP-1不表达。特异性抑制剂PD98059和U0126对丝裂原活化蛋白激酶途径的抑制以及LY294002对磷脂酰肌醇3-激酶的抑制，均强烈抑制Epo诱导的TIMP-1表达和分泌。在没有Epo的情况下，基质金属蛋白酶-9（MMP-9）的潜伏形式和活性形式均分泌到培养基中。在Epo刺激后，MMP-9和前MMP-9的分泌以剂量依赖的方式受到抑制，这与TIMP-1的诱导呈平行关系。在有Epo存在的情况下添加PD98059、U0126和LY294002可恢复UT-7和CD36+细胞中MMP-9的产生。我们的研究结果强烈表明，Epo通过丝裂原活化蛋白激酶和磷脂酰肌醇3-激酶途径对TIMP-1基因和MMP-9的产生进行反向协调调节。
