Kameda H, Ishigami H, Suzuki M, Abe T, Takeuchi T
Division of Rheumatology/Clinical Immunology, and Department of Orthopedics, Department of Internal Medicine, Saitama Medical Center, 1981 Tsujido-cho, Kamoda, Kawagoe, Saitama 350-8550, Japan.
Clin Exp Immunol. 2006 May;144(2):335-41. doi: 10.1111/j.1365-2249.2006.03067.x.
Receptors for platelet-derived growth factor (PDGF) are abundantly expressed on synovial fibroblast-like (SFL) cells from patients with rheumatoid arthritis (RA), and stimulation with PDGF enhances both the anchorage-dependent and -independent growth of RA-SFL cells. To elucidate the molecular mechanisms responsible for the excessive growth of RA-SFL cells and to seek a novel molecular-targeting therapy for RA, we examined the expression of adapter proteins and the effect of the specific inhibition of PDGF receptor activation by imatinib mesylate. Cultured SFL cells were used in the present study after 2-5 passages. The anchorage-dependent and -independent growth patterns of the SFL cells were evaluated using a tetrazolium-based assay and colony formation in 0.3% agar, respectively. Adapter proteins Gab1 and Gab2 were expressed in RA-SFL cells, and both proteins were rapidly (< 1 min) tyrosine-phosphorylated after the stimulation of RA-SFL cells with 10 ng/ml of PDGF and, to a lesser extent, after stimulation with 100 ng/ml of epidermal growth factor (EGF). The inhibition of PDGF receptor tyrosine kinase activation by 1 microM or less of imatinib mesylate specifically suppressed the PDGF-dependent, but not EGF-dependent, tyrosine phosphorylation of various proteins. Moreover, imatinib mesylate abolished both the anchorage-dependent and -independent proliferation of RA-SFL cells induced by PDGF stimulation. These results suggest that Gab adapter proteins are expressed and likely to be involved in the growth signalling of rheumatoid synovial cells and that imatinib mesylate, a key drug in the treatment of chronic myeloid leukaemia, may also be effective for the treatment of RA.
血小板衍生生长因子(PDGF)受体在类风湿关节炎(RA)患者的滑膜成纤维样(SFL)细胞中大量表达,用PDGF刺激可增强RA - SFL细胞的贴壁依赖性和非贴壁依赖性生长。为了阐明导致RA - SFL细胞过度生长的分子机制并寻找针对RA的新型分子靶向治疗方法,我们检测了衔接蛋白的表达以及甲磺酸伊马替尼对PDGF受体激活的特异性抑制作用。本研究使用传代2 - 5次后的培养SFL细胞。分别采用基于四氮唑的检测方法和在0.3%琼脂中进行集落形成实验来评估SFL细胞的贴壁依赖性和非贴壁依赖性生长模式。衔接蛋白Gab1和Gab2在RA - SFL细胞中表达,在用10 ng/ml PDGF刺激RA - SFL细胞后,这两种蛋白均迅速(<1分钟)发生酪氨酸磷酸化,在用100 ng/ml表皮生长因子(EGF)刺激后,磷酸化程度较低。1 μM或更低浓度的甲磺酸伊马替尼抑制PDGF受体酪氨酸激酶激活可特异性抑制各种蛋白的PDGF依赖性酪氨酸磷酸化,但不抑制EGF依赖性酪氨酸磷酸化。此外,甲磺酸伊马替尼消除了PDGF刺激诱导的RA - SFL细胞的贴壁依赖性和非贴壁依赖性增殖。这些结果表明,Gab衔接蛋白表达并可能参与类风湿滑膜细胞的生长信号传导,并且甲磺酸伊马替尼作为治疗慢性髓性白血病的关键药物,可能对RA治疗也有效。