甘草提取物（Gly34）对Hela细胞体外促凋亡作用研究

目的观察甘草提取物（Gly34）对Hela细胞的体外促凋亡作用,为Gly34在抗肿瘤方面的机制研究及应用提供理论依据。方法应用MTT法测定Gly34对Hela细胞增殖的抑制作用;采用AO-EB染色、透射电镜、DNA Ladder实验、流式细胞仪检测其促凋亡作用及对细胞周期的影响。结果 Gly34对Hela细胞体外增殖有明显抑制作用;AO染色表明药物组细胞有明显凋亡迹象,并伴少量细胞坏死现象,而在阴性对照组中细胞形态完好;透射电镜表明Hela细胞在Gly34处理后出现典型凋亡细胞形态改变;DNA Ladder实验发现Hela细胞在Gly34处理后发生DNA片段化断裂,表现为＂梯形＂DNA条带;流式细胞仪（FCM）分析Hela细胞在Gly34处理前后细胞周期均呈现明显的差异,并且药物组细胞表现为G0/G1期阻滞,其中溶媒对照组G0/G1期峰面积为（44.62±0.6）%,1.56μg·mL-1、3.125μg·mL-1、6.25μg·mL^-1药物组G0/G1期峰面积为（53.18±0.7）%、（55.40±0.8）%、（58.67±0.6）%,各组间差异有统计学意义（P〈0.05）;FCM分析Hela细胞在Gly34处理前后细胞凋亡率发生改变,其中溶媒对照组为（0.51±0.6）%,1.56μg·mL^-1、3.125μg·mL^-1、6.25μg·mL^-1药物组分别为（1.25±0.7）%、（21.38±0.8）%、（62.08±0.7）%,各组间差异有统计学意义（P〈0.05）。结论甘草提取物（Gly34）能明显抑制Hela细胞增殖并诱导其发生凋亡,具有抗肿瘤作用。

Role of licorice Anti-tumor Component (Gly34) in apoptosis of Hela cells in vitro

Objective To observe the apoptosis of hela cells induced in vitro by Gly34 which was extracted from glycyrrhiza, and investigate the relevant molecular mechanism. Methods Macroporous resin and Thin-layer chromatography （TLC） were used to get the Gly34 from glycyrrhiza, and the effect of Gly34 on proliferation of hela cells were determined by MTT method. The Gly34-inducing apoptosis in hela was observed by AO/EB fluorescent staining, transmission electron microscopy （TEM）, DNA agarose electrophoresis, and flow cytometry（FCM）. Results Gl34 showed time-dose-dependent inhibitory effect on the proliferation of hela cells. When Gly34-treated cells were stained with AO, apoptosis was observed and there were also a few of necrosis cells; Gly34 also caused DNA ladder formation in a dose-dependent manner; TEM showed that the Gly34-treated-cells were typical apoptosis. G0/G1 phase by FCM showed significant differences between the experimental groups and the control, and the control group was （44.62±0.6）%, and the experimental groups were （53.18±0.7）%, （55.40±0.8）%, （58.67±0.6）% （P〈0.05）. The apoptosis rate showed significant differences between the experimental groups and the control group. The control group was （0.51±0.6）%, and the experimental groups were （1.25±0.7）%, （21.38±0.8）%, （62.08±0.7）% （P〈0.05）. Conclusions Gly34 has the effects of inhabiting growth and inducing apoptosis of hela cells, and had the anti-cancer effect.