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目的:研究鱼藤素对胃癌细胞增殖及凋亡的影响。方法:利用CCK8检测不同浓度(0、1、10和25μmol/L)鱼藤素作用于胃癌细胞MKN-45 24、48、72和96 h后对其增殖的影响;AnnexinⅤ-FITC/PI检测细胞凋亡;流式细胞仪检测细胞周期;利用倒置相差显微镜观察鱼藤素作用后细胞形态的变化;DAPI及Hoechst染色观察细胞核变化。结果:1μmol/L鱼藤素作用于MKN-45细胞24 h后抑制率为(28.82±0.002)%。随着鱼藤素浓度(10、25μmol/L)增加,作用时间(48、72、96 h)延长,其抑制作用增加,表明其抑制作用呈时间-剂量依赖关系。不同浓度(1、10和25μmol/L)鱼藤素作用于细胞48 h后,早期凋亡率、晚期凋亡率和总凋亡率显示鱼藤素显著促进细胞凋亡。细胞周期检测发现,与对照组比较,1μmol/L鱼藤素作用于细胞48 h后,S期细胞比例升高,G_0/G_1期及G_2/M期细胞比例下降;10μmol/L鱼藤素作用于细胞48 h后,G_2/M期细胞比例升高,G_0/G_1期细胞比例下降(P<0.01)。倒置相差显微镜检查镜下则显示,1、10和25μmol/L鱼藤素作用MKN-45细胞48 h后,可见典型凋亡样改变。DAPI及Hoechst染色可见鱼藤素组细胞核皱缩变小、染色质浓聚和典型的凋亡小体。结论:鱼藤素可明显抑制胃癌细胞的增殖、促进凋亡。机制可能与细胞周期阻滞于S期及G_2/M期有关。
Objective: To study the effect of deguelin on proliferation and apoptosis of gastric cancer cells. Methods: CCK8 was used to detect the effect of different concentrations (0, 1, 10 and 25 μmol / L) of deguelin on the proliferation of gastric cancer cells MKN-45 at 24,48,72 and 96 h. AnnexinⅤ-FITC / The cell cycle was detected by flow cytometry. The morphology of cells was observed by inverted phase contrast microscope. The changes of cell nuclear were observed by DAPI and Hoechst staining. Results: The inhibitory rate of MKN-45 cells treated with 1 μmol / L deguelin for 24 h was (28.82 ± 0.002)%. With the increase of deguelin concentration (10,25μmol / L), the action time (48,72,96 h) was prolonged, and the inhibitory effect was increased. It showed that the inhibitory effect was time-dose dependent. At different concentrations (1, 10 and 25 μmol / L) of deguelin for 48 h, the early apoptosis rate, the late apoptosis rate and the total apoptosis rate showed that deguelin significantly promoted cell apoptosis. Cell cycle analysis showed that the percentage of cells in S phase increased and the percentage of cells in G_0 / G_1 phase and G_2 / M phase decreased when treated with 1 μmol / L deguelin for 48 h compared with control group. The effect of 10 μmol / L deguelin After 48 h, the proportion of cells in G 2 / M phase increased and the percentage of cells in G 0 / G 1 phase decreased (P <0.01). Inverted phase contrast microscopy showed that typical apoptosis-like changes were observed in MKN-45 cells treated with 1, 10 and 25 μmol / L deguelin for 48 h. DAPI and Hoechst staining showed degranulation, chromatin condensation and typical apoptotic bodies in deguelin group. Conclusion: deguelin can significantly inhibit the proliferation of gastric cancer cells and promote apoptosis. Mechanisms may be related to cell cycle arrest in S phase and G 2 / M phase.