摘要
目的 通过改变碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor ,bFGF)作用的时间 ,研究bFGF调节鼻咽癌上皮样细胞系 (carcinomaepithelioidcelllines Ⅰ ,CNE Ⅰ )鼻咽癌细胞生长过程与磷脂酰肌醇 3 激酶 /蛋白激酶B (phosphatidylinositol3 kinase/proteinkinaseB ,PI 3 K/PKB)信号转导途径的关系。方法 采用3 2 P掺入法检测细胞质和细胞膜PKB活性。采用WesternBlot检测AP 1转录因子组成成员c fos的蛋白表达水平。结果 当采用外源性bFGF(5 0ng/ml)以不同时间作用于CNE Ⅰ细胞时 ,可提高细胞质和细胞膜PKB活性 ,而PI 3 K特异性抑制剂渥曼青霉素(Wortmannin)预处理 1h后再施加bFGF ,可显著降低细胞质和细胞膜PKB活性。统计学分析 (单因素方差分析 )表明Wortmannin预处理组和未做预处理组之间差异存在显著性 (P <0 0 5 )。当外源性bFGF(5 0ng/ml)以不同时间作用于CNE Ⅰ细胞时 ,c fos表达增加 ,在 30min时达到高峰 ,用Wortmannin(10 0nmol/L)预处理 1h后施加bFGF ,c fos表达与未做预处理组相比有所减少 ,在 30min时间点降低 2 0 % ,在 4 5min时间点降低 35 %。结论 PI3 K、PKB介导bFGF对CNE Ⅰ鼻咽癌细胞的信号转导 ,PKB位于PI3 K下游 ;bFGF通过PI 3 K/PKB通路诱导CNE Ⅰ鼻咽癌细胞c fos表达?
Objective The relationship between basic fibroblast growth factor(bFGF) and phosphatidyl inositol 3-kinase/protein kinase B(PI 3-K/PKB) signal pathway was studied du ring the course of bFGF regulating carcinoma epithelioid cell lines-Ⅰ(CNE-Ⅰ ) nasopharyngeal carcinoma cell proliferation through altering time of bFGF tre atment. Methods PKB activities were measured by advanced Takai method. C-fos expression was ex amined by Western Blot. Results bFGF(50 ng/ml) signific antly stimulated PKB activity,After pretreatment of Wortmannin for 1 h, the PKB activity decreases to the basal levels(P<0.05),compared with control grou p. After treatment of bFGF(50 ng/ml) at various time, the expression of c-fos is increased. The expression of c-fos increases to the peak when the time of bFGF treatment is 30 min. After pretreatment of Wortmannin for 1 h, the express ion of c-fos decreases 20% when the time of bFGF treatment was 30 min and when the time of bFGF treatment was 45 min, the expression of c-fos decreases 35%. Conclusions 1. PI 3-K and PKB mediate bFGF-induced sig nal transduction in CNE-Ⅰ nasopharyngeal carcinoma cell line; PKB lies in downstream of PI 3-K. 2. bFGF induces the increased expression of c-fos thro ugh PI 3-K/PKB in CNE-Ⅰ nasopharyngeal carcinoma cell line.
出处
《中华耳鼻咽喉科杂志》
CSCD
北大核心
2004年第11期679-682,共4页
Chinese Journal of Otorhinolaryngology
