Bodo M, Baroni T, Carinci F, Becchetti E, Bellucci C, Conte C, Pezzetti F, Evangelisti R, Tognon M, Carinci P
Dipartimento di Morfologia ed Embriologia, Università di Ferrara, Italy.
Eur J Cell Biol. 1999 May;78(5):323-30. doi: 10.1016/S0171-9335(99)80066-1.
Bone development is controlled by the autocrine and/or paracrine effects of regulatory molecules. We previously showed that the phenotype of fibroblasts obtained from patients affected by Crouzon's syndrome, an autosomal dominant disease characterized by pathological skull bone development, differed from that of normal cells and was regulated by interleukin treatments. The changes in the relative concentrations of extracellular macromolecules (glycosaminoglycans-GAG, collagen and fibronectin) were associated with abnormal interleukin secretion that affected the microenvironment where the osteogenic processes take place. Mutations in human fibroblast growth factor receptors are now thought to be involved in Crouzon's syndrome. Since coactivation of interleukins and basic fibroblast growth factor (bFGF) is probably implicated in morphogenetic and osteogenic processes and heparan sulphate proteoglycans have a critical role in regulating bFGF activity, the phenotypes of normal and Crouzon osteoblasts were studied and the effects of bFGF on the expression of bFGF, procollagen alpha1 (I), and proteoglycan (PG) genes for biglycan, decorin, betaglycan and syndecan analyzed. Specific human cDNA probes were used to screen the relative levels of mRNA by Northern analysis. Spontaneous or bFGF-modulated release of interleukins was also assayed. The bFGF gene transcript was detected only in Crouzon osteoblasts. We showed for the first time that Crouzon osteoblasts, despite a mutation in the FGF receptor, still responded to exogenous bFGE In fact, the growth factor induced changes in the GAG profile and in the levels of mRNA coding for PG and procollagen alpha1 (I) and down-regulated heparan sulfate GAG chains. ELISA showed that bFGF-induced interleukin secretion differed in normal and Crouzon osteoblasts. The observed differences in PG core protein, procollagen alpha1 (I) and bFGF could be associated with the Crouzon bone phenotype and also should provide further understanding on the molecular basis of the diseased state of bone.
骨发育受调节分子的自分泌和/或旁分泌作用控制。我们先前表明,从患有克鲁宗综合征(一种以病理性颅骨发育为特征的常染色体显性疾病)的患者身上获取的成纤维细胞的表型与正常细胞不同,且受白细胞介素治疗的调节。细胞外大分子(糖胺聚糖 - GAG、胶原蛋白和纤连蛋白)相对浓度的变化与影响成骨过程发生微环境的异常白细胞介素分泌有关。现在认为人类成纤维细胞生长因子受体的突变与克鲁宗综合征有关。由于白细胞介素和碱性成纤维细胞生长因子(bFGF)的共同激活可能参与形态发生和成骨过程,且硫酸乙酰肝素蛋白聚糖在调节bFGF活性中起关键作用,因此研究了正常和成骨不全成骨细胞的表型,并分析了bFGF对bFGF、原胶原α1(I)以及双糖链蛋白聚糖、核心蛋白聚糖、β - 聚糖和多功能蛋白聚糖蛋白聚糖(PG)基因表达的影响。使用特异性人类cDNA探针通过Northern分析筛选mRNA的相对水平。还检测了白细胞介素的自发或bFGF调节释放。仅在成骨不全成骨细胞中检测到bFGF基因转录本。我们首次表明,尽管成骨不全成骨细胞的FGF受体存在突变,但仍对外源bFGF有反应。事实上,生长因子诱导了GAG谱以及编码PG和原胶原α1(I)的mRNA水平的变化,并下调了硫酸乙酰肝素GAG链。ELISA表明,bFGF诱导的白细胞介素分泌在正常和成骨不全成骨细胞中有所不同。观察到的PG核心蛋白、原胶原α1(I)和bFGF的差异可能与成骨不全骨表型有关,也应为进一步了解骨疾病状态的分子基础提供依据。