丹参酮ⅡA诱导人鼻咽癌细胞凋亡及其作用机制的体外实验研究

A Study on Apoptosis of Nasopharyngeal Carcinoma Cell Line Induced by Tanshinone ⅡA and Its Molecular Mechanism

目的 探讨丹参酮 A诱导人鼻咽癌细胞 (CNE- 1)凋亡的作用及其机制。方法 在体外细胞培养的基础上用 0 .5 μg/m l丹参酮 A处理 CNE- 1细胞 4天 ,而后应用光镜、电镜、集落形成试验、DNA凝胶电泳及流式细胞仪分析 CNE- 1细胞增殖及凋亡的改变。结果 经丹参酮 A处理后 ,CNE- 1细胞增殖明显被抑制 ,镜下可见典型的凋亡细胞的特征性改变 ,细胞 DNA呈现“梯状”断裂现象 ,流式细胞仪分析细胞凋亡指数为 16 .9% ,而对照组为 6 .4% ,细胞凋亡相关基因 fas、bax、p5 3及细胞周期负调基因 p2 1表达明显增加 ,bcl- 2表达明显降低。结论 丹参酮 A具有诱导鼻咽癌细胞凋亡的作用 。

Objective To assess effect of TanshinoneⅡA on the apoptosis of human nasopharyngeal carcinoma(NPC)cell line CNE 1 and inquire into the mechanism there in involved. Methods The CNE 1 cells cultured in vitro were treated with 0.5μg/ml Tanshinone ⅡA for 4 days. The morphology of the cells was observed by microscopy. The cell growth and proliferation were measured by cell counting. The DNA break was examined by gel electrophoresis. The cell cycle,apoptosis index (AI) and the expression of apoptosis associated gene were analysed by flow cytometry. Results 0.5μg/ml Tanshinone ⅡA could induce the apoptosis of CNE 1 cells.In the treatment group,the morphologic characteristics of apoptotic cells were observed,the DNA of cells presented 'ladder' break,the cell growth and proliferation were inhibited obviously,and the AI was 16.9% (the AI of control group was 6.4%).The cells were arrested in G 0/G 1 phase and cellular DNA synthesis was inhibited. The expressions of apoptosis associated gene fas,bax,p53 and p21 were up regulated; the expression of bcl 2 was down regulated. Conclusion Tanshinone ⅡA can induce apoptosis of human nasopharyngeal carcinoma cells. Its molecular mechanism may relate to modulation of the apoptosis associated gene expression.