Retinoic acid, GABA-ergic, and TGF-β signaling systems are involved in human cleft palate fibroblast phenotype
Retinoic acid, GABA-ergic, and TGF-β signaling systems are involved in human cleft palate fibroblast phenotype
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DOI：
10.2119/2006-00026.baroni
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发表时间：
2006-09-01
影响因子：
5.7
通讯作者：
Bodo, Maria
中科院分区：
文献类型：
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作者：
Baroni, Tiziano;Bellucci, Catia;Bodo, Maria
在胚胎发生过程中，细胞外基质（ECM）分子，调节分子和生长因子之间的复杂相互作用介导的形态发生过程中参与腭发育。转化生长因子-β（TGF-β）、视黄酸（RA）和γ-氨基丁酸（GABA）能信号系统也可能参与其中。采用（H-3）葡萄糖胺和（S-35）蛋氨酸掺入、阴离子交换色谱、半定量放射性RT-PCR和TGF-β结合试验，我们旨在验证2岁家族性非综合征性唇腭裂和/或腭裂患者（CLP-SP成纤维细胞）和年龄匹配的正常SP（N-SP）成纤维细胞的原代培养物之间存在表型差异。在药理学剂量下，RA会导致许多物种的新生儿腭裂，其影响也被研究。我们发现，与N-SP细胞相比，CLP-SP成纤维细胞合成和分泌更多的糖胺聚糖（GAG）和纤维连接蛋白（FN），ECM的产生发生了变化。在CLP-SP细胞中，TGF-β 3 mRNA表达和TGF-β受体数量增加，RA受体α（RARA）基因表达增加。此外，我们首次证明，GABA受体（GABRB 3）的mRNA表达上调人CLP-SP成纤维细胞。在N-SP和CLP-SP成纤维细胞中，RA减少GAG和FN分泌，增加TGF-β 3 mRNA表达，但减少TGF-β受体的数量。TGF-β受体I型mRNA表达降低，TGF-β受体II型增加，TGF-β受体III型不受影响。RA处理增加RARA基因表达在两个细胞群体，但上调GABRB 3 mRNA表达仅在N-SP细胞。这些结果表明，CLD-SP成纤维细胞与N-SP成纤维细胞相比，在体外表现出异常的表型，并对RA治疗的反应不同，并表明RA，GABA能和TGF-β信号系统之间的改变串扰可能参与人腭裂成纤维细胞表型。
During embryogenesis, a complex interplay between extracellular matrix (ECM) molecules, regulatory molecules, and growth factors mediates morphogenetic processes involved in palatogenesis. Transforming growth factor-beta (TGF-beta), retinoic acid (RA), and gamma-aminobutyric acid (GABA)ergic signaling systems are also potentially involved. Using (H-3)glucosamine and (S-35) methionine incorporation, anion exchange chromatography, semiquantitative radioactive RT-PCR, and a TGF-beta binding assay, we aimed to verify the presence of phenotypic differences between primary cultures of secondary palate (SP) fibroblasts from 2-year-old subjects with familial nonsyndromic cleft lip and/or palate (CLP-SP fibroblasts) and age-matched normal SP (N-SP) fibroblasts. The effects of RA-which, at pharmacologic doses, induces cleft palate in newborns of many species-were also studied. We found an altered ECM production in CLP-SP fibroblasts that synthesized and secreted more glycosaminoglycans (GAGs) and fibronectin (FN) compared with N-SP cells. In CLP-SP cells, TGF-beta 3 mRNA expression and TGF-beta receptor number were higher and RA receptor-alpha (RARA) gene expression was increased. Moreover, we demonstrated for the first time that GABA receptor (GABRB3) mRNA expression was upregulated in human CLP-SP fibroblasts. In N-SP and CLP-SP fibroblasts, RA decreased GAG and FN secretion and increased TGF-beta 3 mRNA expression but reduced the number of TGF-beta receptors. TGF-beta receptor type I mRNA expression was decreased, TGF-beta receptor type II was increased, and TGF-beta receptor type III was not affected. RA treatment increased RARA gene expression in both cell populations but upregulated GABRB3 mRNA expression only in N-SP cells. These results show that CLD-SP fibroblasts compared with N-SP fibroblasts exhibit an abnormal phenotype in vitro and respond differently to RA treatment, and suggest that altered crosstalk between RA, GABAergic, and TGF-P signaling systems could be involved in human cleft palate fibroblast phenotype.