Bcl-2家族蛋白在莱菔硫烷诱导人涎腺腺样囊性癌细胞系ACC-M凋亡中的作用  

Effect of Bcl-2 family on apoptosis of salivary adenoid cystic carcinoma cell line ACC-M induced by sulforaphane

作　　者：贾志宇[1] 郭涛[2] 庄志征 岳磊[1] 杨威[1] 张英怀[1] 

机构地区：[1]河北医科大学第二医院口腔颌面外科,河北石家庄050000 [2]河北省胸科医院口腔科,河北石家庄050041 [3]河北大学附属医院口腔科,河北保定071000

出　　处：《河北医科大学学报》2016年第2期161-165,共5页Journal of Hebei Medical University

基　　金：河北省医学科学研究重点课题(20110340;20130150)

摘　　要：目的观察莱菔硫烷(sulforaphane,SFN)对腺样囊性癌细胞ACC-M中Bcl-2家族成员表达的影响,探讨Bcl-2家族蛋白在SFN诱导ACC-M细胞凋亡中的作用。方法 20或40μmol/L SFN处理ACC-M细胞特定时间后,采用倒置显微镜,Wright-Giemsa染色和透射电镜观察细胞形态学变化,流式细胞术检测细胞凋亡率。40μmol/L SFN处理细胞4、8、16、24h,采用Western blot方法检测Bax、Bak、Bcl-2和Bcl-xL的表达。结果倒置显微镜,Wright-Giemsa染色和透射电镜显示SFN可诱导ACC-M细胞凋亡,流式细胞术检测可见凋亡率随处理时间延长和药物浓度增加而上升,各组间差异有统计学意义(P<0.05)。经40μmol/L SFN处理后,促凋亡蛋白Bax、Bak的表达以及Bax/Bcl-2比值随着时间的延长而增加,抗凋亡蛋白Bcl-2和Bcl-xL的表达随时间延长逐渐降低,差异均有统计学意义(P<0.05)。结论 SFN以浓度-时间依赖方式诱导ACC-M细胞凋亡,并上调Bax和Bak的表达,下调Bcl-2和Bcl-xL的表达。SFN对Bcl-2家族蛋白的调节可能是其诱导凋亡的机制。Objective To study the effect of sulforaphane（SFN）on expression of Bcl-2family in salivary adenoid cystic carcinoma cell line ACC-M,and to evaluate the effect of Bcl-2family on apoptosis of ACC-M cells induced by SFN.Methods After ACC-M cells were treated with 20 or 40μmol/L SFN for desired time,morphology changes of ACC-M cells were observed with inversion phase contrast microscope,Giemsa staining and transmission electron microscope（TEM）.Flow cytometry with Annexin-V-FITC/propidium iodide double staining were used to detect the apoptosis rate of ACC-M.After ACC-M cells were treated with 40μmol/L SFN for 4,8,16 and 24hours,the expression of Bax,Bak,Bcl-2and Bcl-xL were detected by Western blot.Results Image data showed that SFN could induce apoptosis of ACC-M cells.Flow cytometry data showed that apoptosis rate was increased with time and concentration of SFN.There were significantly different among each groups（P〈0.05）.Expression of proapoptosis protein Bax and Bak as well as ratio of Bax/Bcl-2 were increased with treating time,whereas that of antiapoptosis protein Bcl-2and Bcl-xL were decreased by treatment with 40μmol/L SFN for 4,8,16 and 24h.There were significantly different among each groups（P〈0.05）.Conclusion SFN could induce apoptosis of ACC-M cells with time-and concentration-dependent manners,and upregulate expressions of Bax and Bak,and down-regulate expressions of Bcl-2and Bcl-xL,which might be the mechanisms for the apoptosis of ACC-M induced by SFN.

关 键 词：口腔肿瘤 莱菔硫烷 细胞凋亡 

分 类 号：R599[医药卫生—内科学]