Siffroi-Fernandez Sandrine, Cinaroglu Ayca, Fuhrmann-Panfalone Véronique, Normand Guy, Bugra Kuyas, Sahel José, Hicks David
Laboratoire de Physiopathologie Cellulaire et Moléculaire de la Rétine, Institut National de la Santé et de la Récherche Médicale Université Louis Pasteur U.592, Centre Hospitalier Universitaire Régional, Strasbourg, France.
Arch Ophthalmol. 2005 Mar;123(3):368-76. doi: 10.1001/archopht.123.3.368.
Fibroblast growth factors (FGFs) represent potent effectors and play essential roles in both normal development and many pathological processes. Little is known about their possible implication in retinoblastoma growth. We sought to examine FGF high- and low-affinity receptor (FGFR) expression, activation of FGFR1 by acidic FGF (FGF-1), and proliferative effects on Y79 cells.
Expression of FGFR1 to FGFR4 was screened in Y79 cells by means of immunochemical and reverse transcriptase polymerase chain reaction techniques. Tyrosine phosphorylation of FGFR1 induced by FGF was examined by immunoprecipitation after stimulation with FGF-1 in the presence or absence of heparin. Retinoblastoma proliferation was monitored by radiolabeled thymidine incorporation or a vital dye-based assay, after addition of FGF-1 with or without inclusion of a specific FGFR1 neutralizing antibody or FGFR1 antisense oligonucleotides. Low-affinity heparan sulfate proteoglycan coreceptors were blocked through sodium chlorate or heparinase treatment of Y79 cells.
Y79 retinoblastoma expressed all 4 FGFRs, at both the protein and messenger RNA levels. The FGFR1 was differentially phosphorylated in a time- and heparin-dependent manner by FGF-1. Proliferation of Y79 cells induced by FGF-1 was entirely mediated by FGFR1, since inclusion of specific neutralizing antibodies or antisense oligonucleotides completely prevented tumor cell multiplication. Finally, FGF-1-induced proliferation was dependent on the presence and sulfation of heparan sulfate proteoglycan.
Y79 retinoblastoma expresses all 4 FGFRs, but FGFR1 activation entirely accounts for FGF-1-driven cell proliferation.
These studies demonstrate a role for the FGF-1/FGFR1 pathway in retinoblastoma proliferation, and may contribute to developing therapeutic strategies to limit retinoblastoma growth.
成纤维细胞生长因子（FGFs）是强有力的效应分子，在正常发育和许多病理过程中发挥重要作用。关于它们在视网膜母细胞瘤生长中的潜在影响知之甚少。我们试图检测FGF高亲和力和低亲和力受体（FGFR）的表达、酸性FGF（FGF-1）对FGFR1的激活以及对Y79细胞的增殖作用。
通过免疫化学和逆转录聚合酶链反应技术在Y79细胞中筛选FGFR1至FGFR4的表达。在用FGF-1刺激（存在或不存在肝素）后，通过免疫沉淀检测FGF诱导的FGFR1酪氨酸磷酸化。在添加FGF-1（包含或不包含特异性FGFR1中和抗体或FGFR1反义寡核苷酸）后，通过放射性标记的胸腺嘧啶掺入或基于活性染料的检测来监测视网膜母细胞瘤的增殖。通过对Y79细胞进行氯酸钠或肝素酶处理来阻断低亲和力硫酸乙酰肝素蛋白聚糖共受体。
Y79视网膜母细胞瘤在蛋白质和信使RNA水平均表达所有4种FGFR。FGFR1被FGF-1以时间和肝素依赖性方式差异磷酸化。FGF-1诱导的Y79细胞增殖完全由FGFR1介导，因为加入特异性中和抗体或反义寡核苷酸可完全阻止肿瘤细胞增殖。最后，FGF-1诱导的增殖依赖于硫酸乙酰肝素蛋白聚糖的存在和硫酸化。
Y79视网膜母细胞瘤表达所有4种FGFR，但FGFR1的激活完全解释了FGF-1驱动的细胞增殖。
这些研究证明了FGF-1/FGFR1途径在视网膜母细胞瘤增殖中的作用，并可能有助于制定限制视网膜母细胞瘤生长的治疗策略。
