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目的:研究甘草次酸对人食管癌细胞生长的抑制作用及对细胞凋亡的影响。方法:Eca9706细胞接种于96孔板,每孔104个细胞/100μL,培养24 h贴壁,分别加入含不同浓度甘草次酸的培养液100μL,质量浓度分别为95,114,138,166,200 mg·L-1,另设对照孔(加培养液100μL和与溶解药物等量的DMSO),每组设8个复孔。细胞培养48 h,MTT法检测甘草次酸对人食管癌细胞Eca9706增殖的抑制,用流式细胞仪和激光共聚焦显微镜检测细胞凋亡情况。结果:在95~200 mg·L-1质量浓度甘草次酸能有效抑制Eca9706细胞增殖,且呈剂量依赖性,(P<0.05)。83,108,133 mg·L-1的甘草次酸作用Eca9706细胞48 h,细胞凋亡率明显增加,与对照组相比,差异有统计学意义(P<0.01)。激光共聚焦显微镜下对照组细胞膜绿色荧光,以早期凋亡为主,甘草次酸高、低剂量组细胞膜绿色荧光、核红色荧光较多,既有早期凋亡,又有晚期凋亡。结论:甘草次酸可能通过诱导细胞凋亡对食管癌Eca9706细胞增殖起抑制作用。
Objective: To study the inhibitory effect of glycyrrhetinic acid on the growth of human esophageal cancer cells and its effect on apoptosis. Methods: Eca9706 cells were seeded in 96-well plates at a rate of 104 cells / 100μL per well. After culturing for 24 h, 100μL of culture medium containing different concentrations of glycyrrhetinic acid were respectively added into the culture medium. The concentrations of Eca9706 were 95,114,138,166,200 mg · L -1, Control wells (plus 100 μL of culture medium and DMSO equivalent to dissolved drug), each with 8 replicate wells. The cells were cultured for 48 h. The inhibition of glycyrrhetinic acid on the proliferation of human esophageal carcinoma cell line Eca9706 was detected by MTT assay. The apoptosis of cells was detected by flow cytometry and laser confocal microscopy. Results: Glycyrrhetinic acid (95-200 mg · L-1) could inhibit the proliferation of Eca9706 cells in a dose-dependent manner (P <0.05). The apoptosis rate of Eca9706 cells treated with glycyrrhetic acid at doses of 83,108 and 133 mg · L-1 for 48 h significantly increased compared with the control group (P <0.01). Under the confocal laser scanning microscope, the cell membrane of the control group was green fluorescence, which was mainly in the early stage of apoptosis. The glycyrrhetinic acid high and low dose groups had green fluorescence and nuclear red fluorescence, both early and late apoptosis. Conclusion: Glycyrrhetinic acid may inhibit the proliferation of esophageal cancer Eca9706 cells by inducing apoptosis.