微管不稳定蛋白stathmin基因沉默影响鼻咽癌5-8F细胞系增殖与凋亡

目的: 构建微管不稳定蛋白stathmin基因siRNA质粒表达载体,探讨stathmin沉默对鼻咽癌5-8F细胞的增殖和凋亡作用。方法: 合成的stathmin基因特异性干扰片段单链退火形成双链后,亚克隆于siRNA质粒表达载体 pGenesil-1.1,质粒经鉴定后,采用脂质体将其转入鼻咽癌5-8F细胞;Western blotting检测stathmin表达;MTT实验分析细胞增殖;流式细胞术分析细胞凋亡。结果: Stathmin沉默组5-8F细胞的抑制率为(53.01±1.12)%,与试剂对照组 和阴性对照组 比较,差异显著(P<0.05)。Stathmin沉默组5-8F细胞的凋亡率为(8.75±0.67)%,与试剂对照组 和阴性对照组 比较,差异显著(P<0.05)。结论: Stathmin沉默抑制鼻咽癌5-8F细胞增殖,诱导鼻咽癌5-8F细胞凋亡。

Effect of microtubule-destabilizing protein stathmin gene silencing on proliferation and apoptosis of nasopharyngeal carcinoma cell line 5-8F

AIM: To investigate the effects of stathmin gene silencing on nasopharyngeal carcinoma cell line 5-8F.METHODS: Double-strand siRNA targeting to stathmin gene was obtained by chemical synthesis and annealing,and was sub-cloned into the vector pGenesil-1.1.The plasmid was introduced into 5-8F cells by liposome-mediated transfection.The gene expression of stathmin,and the proliferation,morphology and apoptosis of the cells were analyzed by Western blotting,MTT assay and flow cytometry.RESULTS: The cell suppression rate in stathmin gene silencing group was(53.01±1.12)%,significantly higher than that in transfection reagent group (13.43±0.55)%] and in negative control group (15.57±0.94)%].The cell apoptotic rate in stathmin gene silencing group was(8.75±0.67)%,also significantly higher than that in transfection reagent group (1.56±0.13)%] and in negative control group (1.90±0.47)%](P<0.05).CONCLUSION: Silencing of stathmin gene in nasopharyngeal carcinoma cells inhibits the cell proliferation and induces cell apoptosis.