染氟成骨细胞差异表达基因的cDNA克隆

Cloning of differentially displayed cDNAs involved in NaF-treated osteoblasts

目的 探讨过量氟对成骨细胞基因表达的影响。方法 采用mRNA差异显示聚合酶链反应 (DD PCR)技术 ,对染氟 2周 (氟化钠 2 .0mmol/L)的成骨细胞与正常成骨细胞之间基因的差异表达进行比较分析。结果 在已克隆到的 6个差异表达的基因片段中 ,大鼠核糖体蛋白L5基因、大鼠Na+ K+ ATP酶 β3基因、大鼠细胞核定位信号蛋白质受体α2基因和大鼠高尔基体 p2 8基因在染氟成骨细胞中低表达 ,小鼠遍在蛋白缀合酶基因及我们发现的 1个新基因片段在染氟成骨细胞中表达上调。结论 成骨细胞染氟后基因表达发生改变 ,这些基因多与蛋白质的合成、转运及加工有关 ,提示过量氟可通过改变这些基因的表达而影响成骨细胞的蛋白质合成。另发现 1个与过量氟相关的新基因。

Objective To explore the effects of excessive fluoride on the gene expression of rat osteoblasts. Methods Rat osteoblasts were treated with 2.0 mmol/L of sodium fluoride (NaF) for two weeks in vitro, and difference in the gene expression between the NaF-treated and normal osteoblasts was compared with mRNA differential display (DD-PCR) technique. Results Among the six differentially expressed gene fragments which had been cloned, expression of the ribosomal protein L5 gene, ATPase Na +K + transporting beta polypeptide 3 gene, karyopherin alpha 2 gene and cis-Golgi body p28 gene was lower and expression of ubiquitous-conjugating enzyme E2D 3 gene and a newly-discovered gene fragment in this study showed up-regulated in the NaF-treated osteoblasts of the rats.Conclusions Expression of genes changed in the osteoblasts after treatment with fluoride for two weeks and most of them associated with synthesis, transportation and processing of protein.It suggested that excessive fluoride could affect the protein synthesis in osteoblasts by changing the expression of the related genes.A novel gene related to excessive fluoride exposure was also found.