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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 effect of Gly34 on the apoptosis of Hela cells in vitro and to provide a theoretical basis for the study on the mechanism of Gly34 in antitumor and its application. Methods The inhibitory effect of Gly34 on the proliferation of Hela cells was determined by MTT assay. The effects of Gly34 on apoptosis were detected by AO-EB staining, transmission electron microscopy, DNA Ladder and flow cytometry. Results Gly34 significantly inhibited the proliferation of Hela cells in vitro. The results of AO staining showed that the cells in the drug group showed obvious signs of apoptosis accompanied with a small amount of cell necrosis while the cells in the negative control group were intact. Transmission electron microscopy showed that HeLa cells treated with Gly34 DNA ladder showed that DNA fragmentation was detected in Hela cells after treated with Gly34, which was characterized by “trapezoidal” DNA bands. Flow cytometry (FCM) analysis of Hela cells before and after Gly34 treatment (P <0.05). The G0 / G1 phase arrest was observed in the drug-treated group, with the peak areas of G0 / G1 phase in the vehicle control group being (44.62 ± 0.6)%, 1.56μg · mL-1 and 1.125μg · mL- The peak area of G0 / G1 phase was (53.18 ± 0.7)%, (55.40 ± 0.8)% and (58.67 ± 0.6)% respectively in the group of 1, 6.25 μg · mL-1 and the difference was statistically significant ); FCM analysis of Hela cells in the Gly34 treatment before and after the change in the rate of apoptosis, the vehicle control group was (0.51 ± 0.6)%, 1.56μg · mL-1 · 3.125μg · mL-1 · 6.25μg · mL- (1.25 ± 0.7)%, (21.38 ± 0.8)% and (62.08 ± 0.7)%, respectively. There was significant difference among the groups (P <0.05). Conclusion Gly34 extract can significantly inhibit the proliferation of Hela cells and induce its apoptosis.