摘要
目的:初步探讨以小分子干扰RNA沉默碱性成纤维细胞生长因子(bFGF)诱导胶质瘤细胞系U251凋亡的机制。方法:将U251细胞分为正常对照组、空载体组和实验组,按4×105个细胞每孔接种于6孔细胞培养板,培养24h后,空载体组和实验组按MOI=50分别进行空载体腺病毒(rAd5-null)和含有bFGF小分子干扰RNA(siRNA)的重组腺病毒(rAd5-bFGF)转染,转染72h后检测各组细胞的凋亡及细胞周期变化情况及其相关蛋白的表达。结果:与正常对照组、空载体组比较,实验组U251细胞经转染bFGF-siRNA后出现了明显的细胞凋亡(P<0.05),但细胞周期未见变化(P>0.05);Westernblot结果显示,实验组U251胶质瘤细胞经转染bFGF-siRNA72h后,bFGF蛋白表达明显降低,同时STAT3及Bcl-2蛋白表达降低,Bax及Caspase-3蛋白表达增强。结论:bFGF小分子干扰RNA能够诱导U251细胞凋亡,其作用可能是通过调节STAT3信号转导通路实现的。
Objective： To preliminarily investigate the mechanism of small interfering RNA （siRNA） induced apoptosis in glioma U251 cells by silencing basic fibroblast growth factor （bFGF）. Methods： The U251 ceils were divided into the normal control group, mock group and experiment group. Each group was cultured in 6-well cell culture plates with 4x10s cells per well. After 24 hours of culture, the mock and experiment groups were transfected with mock vector （rAd5-null） and the recom- binant adenovirus carrying bFGF-siRNA （rAdS-bFGF） respectively at a multiplicity of infection （MO[） of 50. The apoptosis and the change of cell cycle were detected by flow cytometry 72 hours after transfection. The expression of related proteins was revealed by the method of Western blot. Results： After U251 cells were transfected with bFGF-siRNA, the apoptosis effect was observed obviously （P 〈 0.05）, but showed no difference in cell cycle distribution （P 〉 0.05） in experiment group compared with those of normal control group and mock group. The results of Western blot showed that after 72 hours of transfection the bFGF protein decreased obviously in the experiment group. Meanwhile, the expressions of Bcl-2 and the signal transducers and activators of transcription 3 （STAT3） proteins decreased while the expressions of Bax and Caspase-3 proteins increased in the experiment group. Conclusion： The bFGF siRNA can induce apoptosis in U251 cells and this effect may be achieved by regulating STAT3 signal pathway.
出处
《天津医药》
CAS
北大核心
2009年第5期337-339,共3页
Tianjin Medical Journal
基金
国家自然科学基金资助项目(项目编号:30672158)
