Zhang Fang, Li Cong, Halfter Hartmut, Liu Jingwen
Department of Veterans Affairs Palo Alto Health Care System, CA 94304, USA.
Oncogene. 2003 Feb 13;22(6):894-905. doi: 10.1038/sj.onc.1206158.
A number of studies have demonstrated that the STAT pathway is an important signaling cascade utilized by the IL-6 cytokine family to regulate a variety of cell functions. However, the downstream target genes of STAT activation that mediate the cytokine-induced cellular responses are largely uncharacterized. The aims of the current study are to determine whether the STAT signaling pathway is critically involved in the oncostatin M (OM)-induced growth inhibition and morphological changes of MCF-7 cells and to identify STAT3-target genes that are utilized by OM to regulate cell growth and morphology. We show that expression of a dominant negative (DN) mutant of STAT3 in MCF-7 cells completely eliminated the antiproliferative activity of OM, whereas expression of DN STAT1 had no effect. The growth inhibition of breast cancer cells was achieved through a concerted action of OM on cell cycle components. We have identified four cell cycle regulators including c-myc, cyclin D1, c/EBPdelta, and p53 as downstream effectors of the OM-activated STAT3 signaling cascade. The expression of these genes is differentially regulated by OM in MCF-7 cells, but is unaffected by OM in MCF-7-dnStat3 stable clones. We also demonstrate that the OM-induced morphological changes are correlated with increased cell motility in a STAT3-dependent manner. Expression analysis of extracellular matrix (ECM) proteins leads to the identification of fibronectin as a novel OM-regulated ECM component. Our studies further reveal that STAT3 plays a key role in the robust induction of fibronectin expression by OM in MCF-7 and T47D cells. These new findings provide a molecular basis for the mechanistic understanding of the effects of OM on cell growth and migration.
多项研究表明,STAT信号通路是白细胞介素-6细胞因子家族用于调节多种细胞功能的重要信号级联反应。然而,介导细胞因子诱导的细胞反应的STAT激活的下游靶基因在很大程度上尚未明确。本研究的目的是确定STAT信号通路是否在抑瘤素M(OM)诱导的MCF-7细胞生长抑制和形态变化中起关键作用,并鉴定OM用于调节细胞生长和形态的STAT3靶基因。我们发现,在MCF-7细胞中表达STAT3的显性负性(DN)突变体完全消除了OM的抗增殖活性,而DN STAT1的表达则没有影响。乳腺癌细胞的生长抑制是通过OM对细胞周期成分的协同作用实现的。我们已经鉴定出四种细胞周期调节因子,包括c-myc、细胞周期蛋白D1、c/EBPδ和p53,作为OM激活的STAT3信号级联反应的下游效应物。这些基因的表达在MCF-7细胞中受到OM的差异调节,但在MCF-7-dnStat3稳定克隆中不受OM的影响。我们还证明,OM诱导的形态变化与细胞运动性增加以STAT3依赖的方式相关。细胞外基质(ECM)蛋白的表达分析导致鉴定纤连蛋白为一种新的OM调节的ECM成分。我们的研究进一步揭示,STAT3在OM对MCF-7和T47D细胞中纤连蛋白表达的强烈诱导中起关键作用。这些新发现为从机制上理解OM对细胞生长和迁移的影响提供了分子基础。