大蒜素对人喉癌Hep-2细胞增殖、凋亡的影响

Effects of Allitridi on human laryngeal squamous cell carcinoma Hep-2 cell growth and its mechanism

目的研究大蒜素(Allitridi)对人喉癌Hep-2细胞增殖、凋亡的影响,并探讨其作用机制。方法分别以不同浓度大蒜素(25、50、100 mg·L^(-1))和顺铂(1.8 mg·L^(-1))干预对数生长期人喉癌Hep-2细胞,药物干预24 h后经Hoechst染色后利用倒置显微镜观察细胞的形态,采用MTT比色法测定大蒜素对Hep-2细胞的增殖抑制作用,采用FCM测定细胞周期分布、检测细胞凋亡率;采用RT-PCR检测Bax、Bcl-2 mRNA表达水平,Western-blot检测caspase-3蛋白表达。结果大蒜素各组和顺铂1.8mg·L^(-1)组人喉癌Hep-2细胞较空白对照组出现不同程度损伤,以大蒜素100 mg·L^(-1)组损伤最为严重;大蒜素(50、100 mg·L^(-1))组和顺铂1.8 mg·L^(-1)组人喉癌Hep-2细胞增殖抑制率显著升高,细胞周期G0/G1期显著增长、G2/M期显著缩短,细胞凋亡数量明显增多、凋亡率显著升高,凋亡相关基因bcl-2 mRNA表达显著下调而Bax mRNA显著上调、Bax/bcl-2表达比值显著升高,促凋亡蛋白caspase-3表达显著上调,上述均差异有统计学意义(P<0.05,P<0.01)。结论大蒜素对人喉癌Hep-2细胞生长具有抑制作用,其机制可能与大蒜素能够阻滞人喉癌Hep-2细胞周期进程、抑制细胞增殖并促进其凋亡有关。

Objective To investigate the effects of Allitridi on human laryngeal squamous cell carcinoma Hep-2 cell growth and its mechanism.Methods The laryngeal squamous Hep-2 cells in logarithmic growth phase were treated by Allitridi （25,50,100 mg·L-1） and Cisplatin （1.8 mg·L-1）.Twenty-four hours after the drugs were given,the growth status of Hep-2 cells were observed by inverted microscope after Hoechst staining,the cellular growth inhibition rate was calculated by MTT,cell cycle and apoptosis rate were analyzed by flow cytometry,the expressions of Bax mRNA and Bcl-2 mRNA were detected by RT-PCR,and the expression of caspase-3 protein was detected by estern blotting.Results Compared with blank control group,there were different degrees of cellular damage in Allitridi （25,50,100 mg＆#183;L-1） groups and Cisplatin （1.8 mg·L-1） group,among which the damage in Allitridi （100 mg·L-1） group was the severest.The cellular growth inhibition rate of Allitridi （50,100 mg·L-1） groups and Cisplatin （1.8 mg·L-1） group were significantly increased;the cell cycle was extended at G0/G1 phase and shortened at G2/M phase;the apoptosis rate was significantly increased;the expression of Bcl-2 mRNA was significantly decreased,while the expression of Bax mRNA was significantly increased,and the ratio of Bax/Bcl-2 was significantly increased;the expression of caspase-3 protein was significantly increased.All of the above-mentioned differences were significant （P〈0.05,P〈0.01）.Conclusions Allitridi has inhibitive effects on the growth of human laryngeal cancer Hep-2 cells,the mechanism of which is perhaps related to Allitridi′s capacity of altering cell cycle,depressing its proliferation and promoting its apoptosis.