甘草提取物(Gly34)对Hela细胞体外促凋亡作用研究

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

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。

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.