体外刺激对培养成纤维细胞功能与表达c-fos基因的影响

THE STIMULATING EFFECTS OF bFGF ON FIBROBLAST FUNCTION AND ITS C-FOS GENE EXPRESSION

目的 研究采用碱性成纤维细胞生长因子 (b FGF)刺激对体外培养成纤维细胞形态、功能以及表达原癌基因 c- fos的影响 ,探讨成纤维细胞生长因子与原癌基因之间在调控创面愈合中可能的网络机制。方法将处于对数生长期的乳鼠成纤维细胞分成 b FGF刺激组和对照组 ,在分别采用 b FGF和对照物刺激后继续培养 ,于刺激后 1小时和 3、5及 7天采集细胞用于形态学和细胞活力检测。同时用 SP法对原位培养法和甩片法收集的成纤维细胞检测 c- fos基因表达。结果 培养的成纤维细胞经 b FGF刺激后其形态较对照明显增大 ,MTT检测活性明显增高 ,在刺激后表达 c- fos基因显著增加 ,其中以刺激后 1小时最为明显。结论 b FGF刺激可以使培养的成纤维细胞形态与功能发生明显改变 ,使 c- fos基因表达显著增加。结合既往研究表明 ,c- fos不仅可以直接上调b FGF基因表达 ,同时 b FGF本身也可以诱导 c- fos基因表达 ,表明二者之间可能存在相互作用的网络机制。其可能的调控途径涉及 ras以及酪氨酸激酶等。

Objective To study the stimulating effects of basic fibroblast growth factor(bFGF) on fibroblast function and its ability to expression of c fos gene. Furthermore, to explore the possible network action between bFGF and oncogene in modulating wound healing. Methods Cultured rat fibroblasts were divided into bFGF stimulating group and control group. Fibroblasts in bFGF stimulating group were treated with bFGF in a dosage of 40ng/culture hole, while the control fibroblasts were treated with the same vehicle without bFGF. The morphology, cell vitality and their ability to express c fos gene in the fibroblasts in both groups were studied with MTT and immunohistochemical methods. Results All fibroblasts in bFGF treated groups were enlarged and showed increased vitality with MTT method. C fos gene expression in bFGF stimulating group was increased, especially in nucleus when compared with those in control group. Conclusion The results show that the function and the ability to express c fos gene in bFGF treated fibroblasts are enhanced. Combined with our previous studies, it may make a conclusion that there is a network regulation mechanism between growth factors and some oncogenes.