大蒜素对人舌鳞状细胞癌Tca8113细胞凋亡及细胞周期的影响

Effect of Allitridi on apoptosis and cell cycle of human tongue squamous cell carcinoma Tca8113 cells

关键词:  大蒜素  舌鳞状细胞癌  Tca8113细胞  凋亡  细胞周期

Key words:  Allitridi  tongue squamous cell carcinoma  Tca8113 cell  apoptosis  cell cycle

摘要:

[目的]研究大蒜素对人舌鳞状细胞癌Tca8113细胞凋亡以及细胞周期的影响并初探其机制。[方法]取对数生长期Tca8113细胞设空白对照组、大蒜素（50、25、12.5 μg/mL）组和顺铂（40 μg/mL）组，给药48 h后流式细胞术分析细胞周期的改变并检测细胞凋亡状况，逆转录聚合酶链式反应法（RT-PCR）检测B淋巴细胞瘤-2（Bcl-2）mRNA、Bcl-2相关X蛋白（Bax）mRNA表达，蛋白免疫印迹法检测细胞周期相关调控蛋白周期蛋白（cyclin B1、cyclin D1）和周期蛋白依赖性激酶（CDK1、CDK2）表达。[结果]与空白对照组比较，大蒜素（50、25 μg/mL）组细胞周期G₂/M期比例显著提高（P<0.05），细胞凋亡率显著升高（P<0.01），Bcl-2 mRNA表达显著下调且Bax mRNA表达显著上调（P<0.05或P<0.01），Bax/Bcl-2值显著提高（P<0.01），cyclin B1蛋白和CDK1蛋白显著上调、cyclin D1蛋白和CDK2蛋白显著下调（P<0.05或P<0.01）。[结论]大蒜素具有促进人舌鳞状细胞癌Tca8113细胞凋亡、阻滞细胞有丝分裂的作用，其机制可能与大蒜素影响凋亡相关调控基因及细胞周期相关调控蛋白表达有关。

Abstract:

[Objective] To study the effect of Allitridi on apoptosis and cell cycle of human tongue squamous cell carcinoma Tca8113 cells.[Methods] The human tongue squamous cell carcinoma Tca8113 cells in logarithmic growth period were divided randomly into five groups:normal control group, Allitridi (50, 25, 12.5 μg/mL) groups and Cisplatin 40 μg/mL group. 48h after the drugs were given, the cellular growth inhibition rate was detected by MTT, cell cycle and apoptosis rate were analyzed by flow cytometry, the expression of Bax and Bcl-2 mRNA were detected by RT-PCR; the expression of cycle-related regulation of protein (cyclin B1, cyclin D1) and cyclin-dependent kinase (CDK1, CDK2) were detected by Western blotting.[Results] Compared with normal control group. The ratio of the cells in G₂/M phase in Allitridi (50, 25 μg/mL) groups were significantly increased (P<0.05), the apoptosis rate were significantly increased (P<0.01); the expression of Bcl-2 mRNA were significantly decreased, while the expression of Bax mRNA were significantly increased and the ratio of Bax/Bcl-2 were significantly increased (P<0.05, P<0.01); the expression of Cyclin B1 protein and CDK1 protein were significantly up-regulated, the expression of cyclin D1 protein and CDK2 protein were significantly down-regulated (P<0.05, P<0.01).[Conclusion] Allitridi can promote human tongue squamous cell carcinoma Tca8113 cells apoptosis and block cell mitosis, which perhaps related to its effects of altering the expression of apoptosis-related regulatory genes and cell cycle-related regulatory proteins.