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目的:研究葛花中的鸢尾异黄酮对SMMC-7721人肝癌细胞株增殖的抑制作用。方法:将野葛花的甲醇提取物用盐酸加热水解得到葛花总异黄酮,总异黄酮经硅胶柱层析分离获得鸢尾异黄酮。SMMC-7721人肝癌细胞培养至对数生长期后,加入不同浓度鸢尾异黄酮进行干预,在倒置显微镜下观察细胞形态的变化;采用四甲基偶氮唑(MTT,methyl thiazolyl tetrazolium)法分析肝癌细胞生长抑制作用;使用Annexin V-FITC/PI双染色,经流式细胞仪检测鸢尾异黄酮对细胞早期凋亡率的影响。结果:鸢尾异黄酮明显减少肝癌细胞贴壁数量,使癌细胞变小变圆;在1.0~8.0μg.mL-1范围内浓度依赖性地抑制人肝癌细胞SMMC-7721的增殖;鸢尾异黄酮在5.00,10.00和20.00μg.mL-1浓度下均能显著提高肝癌细胞株早期凋亡率(P<0.01)。结论:鸢尾异黄酮对人肝癌细胞SMMC-7721增殖具有明显的抑制作用,其机制可能与促进癌细胞凋亡有关。
Objective: To study the inhibitory effect of iris isoflavones on the proliferation of SMMC-7721 human hepatoma cell line. Methods: The methanol extract of Pueraria lobata was hydrolyzed with hydrochloric acid to obtain the total flavonoids of Pueraria lobata, and the total isoflavones were separated by silica gel column chromatography to obtain iris isoflavones. After the SMMC-7721 human hepatocarcinoma cells were cultured to logarithmic growth phase, different concentrations of isoflavones were intervened to observe the changes of cell morphology under an inverted microscope. The hepatocellular carcinoma (HCC) was analyzed by MTT (methyl thiazolyl tetrazolium) Cell growth inhibition; Annexin V-FITC / PI double staining, flow cytometry test iris isoflavones on cell apoptosis rate. Results: The iris isoflavones significantly reduced the number of hepatocellular carcinoma cells adherent, so that the cancer cells become smaller round; in a concentration range of 1.0 ~ 8.0μg.mL-1 inhibition of human hepatoma SMMC-7721 proliferation; iris Isoflavones in 5.00,10.00 and 20.00μg.mL-1 concentrations can significantly improve the early apoptosis rate of hepatoma cell lines (P <0.01). Conclusion: Isoflavone has significant inhibitory effect on the proliferation of human hepatocellular carcinoma cell line SMMC-7721, which may be related to the promotion of cancer cell apoptosis.