粉防己碱对人鼻咽癌细胞株CNE增殖抑制和凋亡作用的研究

Effects of tetrandrine on proliferation and apoptosis of nasopharyngeal carcinoma cell line CNE

目的:探讨粉防己碱(Tet)对人鼻咽癌细胞株CNE增殖抑制和凋亡的影响。方法:将人鼻咽癌细胞株CNE分为不同浓度Tet处理组,并设立对照组。采用四甲基偶氮唑蓝(MTT)法观察Tet对人鼻咽癌细胞株CNE体外生长的抑制作用;采用透射电镜、琼脂糖凝胶电泳技术、流式细胞术Annenxin V/PI EGFP观察Tet对人鼻咽癌细胞株CNE凋亡的影响;采用逆转录-多聚酶链反应技术(RT-PCR)观察Tet对人鼻咽癌细胞株CNE凋亡相关基因mRNA表达的影响。结果:Tet呈浓度依赖性抑制癌细胞体外生长(P<0.05)。Tet处理细胞72 h后,电镜下可见细胞缩小,核裂解,凋亡小体形成;随着Tet浓度的提高,各Tet处理组DNA凝胶电泳梯形条带和细胞凋亡率渐趋明显。与对照组相比较,Bcl-2 mRNA的表达渐减弱,Bax基因mRNA表达渐增强(P<0.05)。结论:Tet可抑制癌细胞生长,促进癌细胞凋亡,其抗癌作用机制可能与下调Bcl-2基因表达和上调Bax基因表达有密切联系。

Objective ：To investigate the effects of tetrandrine （Tet） on proliferaton and apoptosis of nasopharyngeal carcinoma cell fine CNE. Methods：CNE cells were divided into groups and treated by Tet at different concentrations. Inhibitory effects of tetrandrine were determined by MTT assay. The morphologic changes of CNE cells were observed by transmission electron microscope. DNA fragmentations were determined by gel electrophoresis assay. Cell apoptosis was investigated by flow cytometer. The expressions of apoptosis-related genes were detected by retrotranscriptase polymerase chain reaction （RT- PCR）. Results ：Tetrandrine possessed inhibitory effect on CNE cell proliferation in a concentration-dependent manner （P 〈 0.05）. When treated with Tet for 72 h, cell shrinkage, nuclear fragmentation and apoptotic body formation were observed under transmission electron microscope. With concentration of Tet raised, ladder-shaped straps in DNA gel electrophoresis and apoptosis manifested gradually. Compared with those of control group, expression of Bcl-2 mRNA decreased gradually ; but expression of Bax mRNA increased gradually （ P 〈 0. 05 ）. Conclusion ： Tetrandrine may inhibit growth of cancer cell, induce apoptosis. The anti-cancer mechanism of Tet may be involved in up-regulation of Bax gene expression and down-regulation of Bcl-2 gene expression.