Caspase在莱菔硫烷诱导人腺样囊性癌细胞系ACC-M凋亡中的作用

Effect of caspases on apoptosis induced by SFN in salivary adenoid cystic carcinoma cell line ACC-M in vitro

目的体外研究Caspase-3,8,9在莱菔硫烷(Sulforaphane,SFN)诱导人腺样囊性癌细胞系ACC-M凋亡中的作用,探讨其可能的机制,为临床应用SFN治疗腺样囊性癌提供理论依据。方法 ACC-M细胞经40μmol/L SFN处理4,8,16,24h后,采用分光光度法分别检测Caspase-3,8,9的活性变化。ACC-M细胞采用50μmol/L的Caspase抑制剂Z-VAD-FMK预处理2h后,加入40μmol/L SFN,设阴性和阳性对照,流式细胞术检测细胞凋亡率情况。结果 SFN诱导涎腺腺样囊性癌ACC-M凋亡过程中,Caspase-3,8,9的活性均在4小时时即出现增高,并持续到24小时,且在16小时时活性最高,各时间组与对照组相比均有统计学差异。50μmol/L Caspase抑制剂Z-VAD-FMK预处理细胞2小时,能显著抑制SFN诱导的ACC-M细胞凋亡。结论 SFN在诱导ACC-M细胞凋亡过程中,引起Caspase-3,8,9活性升高,Caspase在SFN诱导ACC-M细胞凋亡中起重要作用。

Objective To evaluate the effect of caspase-3, 8 and 9 on apoptosis induced by Sulforaphane （SFN） in salivary adenoid cystic carcinoma cell line ACC-M in vitro, in order to explore the possible mechanism of SFN- induced apoptosis and provide theoretical evidence for clinical application. Methods After ACC-M cells were treated with 40μmol/L of SFN for 4, 8, 16, 24h respectively,the activities of caspase-3, 8 and 9 were detected with spectrophotometr. 50tLmol/L Caspase inhibitor Z-VAD-FMK was added in the group of SFN＋Caspase inhibitor 2h before treatment at a final concentration of 50μM. And then cells were incubated with 0.04% DMSO, 40μM SFN, or 40μM SFN＋5OμM Caspase inhibitor for 24h. The apoptosis of the ceils was detected with flow cytometry. Results The activities of caspase-3, 8 and 9 on apoptosis induced by SFN in salivary adenoid cystic carcinoma cell line ACC-M obviously increased from 4h to 24h and reach the peak value at 16h. Pretreating with 50μmol/L Caspase inhibitor Z-VAD-FMK for 2h before SFN treatment could significantly inhibit SFN-induced apoptosis. Conclusion Activities of Caspase-3, 8 and 9 were increased significantly and play important roles in SFN-induced apoptosis of ACC-M