Yu J, Deuel T F, Kim H R
Department of Pathology, Barbara Ann Karmanos Cancer Institute, Wayne State University, School of Medicine, Detroit, Michigan 48201, USA.
J Biol Chem. 2000 Jun 23;275(25):19076-82. doi: 10.1074/jbc.M910329199.
Platelet-derived growth factor (PDGF) is a potent mitogen for mesenchymal cells. The PDGF B-chain (c-sis proto-oncogene) homodimer (PDGF BB) and v-sis, its viral counterpart, activate both alpha- and beta-receptor subunits (alpha-PDGFR and beta-PDGFR) and mediate anchorage-independent growth in NIH3T3 cells. In contrast, the PDGF A chain homodimer (PDGF AA) activates alpha-PDGFR only and fails to induce phenotypic transformation. In the present study, we investigated alpha- and beta-PDGFR specific signaling pathways that are responsible for the differences between the transforming ability of PDGF AA and BB. To study PDGF BB activation of beta-PDGFR, we established NIH3T3 clones in which alpha-PDGFR signaling is inhibited by a dominant-negative alpha-PDGFR, or an antisense construct of alpha-PDGFR. Here, we demonstrate that beta-PDGFR activation alone is sufficient for PDGF BB-mediated anchorage-independent cell growth. More importantly, inhibition of alpha-PDGFR signaling enhanced PDGF BB-mediated phenotypic transformation, suggesting that alpha-PDGFR antagonizes beta-PDGFR-induced transformation. While both alpha- and beta-receptors effectively activate ERKs, alpha-PDGFR, but not beta-PDGFR, activates stress-activated protein kinase-1/c-Jun NH(2)-terminal kinase-1 (JNK-1). Inhibition of JNK-1 activity using a dominant-negative JNK-1 mutant markedly enhanced PDGF BB-mediated anchorage-independent cell growth, demonstrating an antagonistic role for JNK-1 in PDGF-induced transformation. Consistently, overexpression of wild-type JNK-1 reduced PDGF BB-mediated transformation. Taken together, the present study showed that alpha- and beta-PDGFRs differentially regulate Ras-mitogen-activated protein kinase pathways critical for regulation of cell transformation, and transformation suppressing activity of alpha-PDGFR involves JNK-1 activation.
血小板衍生生长因子(PDGF)是一种对间充质细胞有强大作用的促有丝分裂原。PDGF B链(c-sis原癌基因)同二聚体(PDGF BB)及其病毒对应物v-sis可激活α和β受体亚基(α-PDGFR和β-PDGFR),并介导NIH3T3细胞的不依赖贴壁生长。相比之下,PDGF A链同二聚体(PDGF AA)仅激活α-PDGFR,且不能诱导表型转化。在本研究中,我们调查了α和β-PDGFR特异性信号通路,这些通路导致了PDGF AA和BB转化能力的差异。为研究PDGF BB对β-PDGFR的激活作用,我们建立了NIH3T3克隆,其中α-PDGFR信号通过显性负性α-PDGFR或α-PDGFR的反义构建体受到抑制。在此,我们证明单独的β-PDGFR激活足以实现PDGF BB介导的不依赖贴壁细胞生长。更重要的是,抑制α-PDGFR信号增强了PDGF BB介导的表型转化,表明α-PDGFR拮抗β-PDGFR诱导的转化。虽然α和β受体都能有效激活细胞外信号调节激酶(ERK),但α-PDGFR而非β-PDGFR能激活应激激活蛋白激酶-1/c-Jun氨基末端激酶-1(JNK-1)。使用显性负性JNK-1突变体抑制JNK-1活性显著增强了PDGF BB介导的不依赖贴壁细胞生长,证明JNK-1在PDGF诱导的转化中起拮抗作用。一致地,野生型JNK-1的过表达降低了PDGF BB介导的转化。综上所述,本研究表明α和β-PDGFR对细胞转化调控至关重要的Ras-丝裂原活化蛋白激酶通路有不同调节作用,且α-PDGFR的转化抑制活性涉及JNK-1激活。