秦皮乙素的制备及对人肝癌细胞SMMC-7721体外增殖的影响
Prepare Esculetin and Study the Effect of Esculetin on Proliferation of Human Heptocellular Carcinoma Cell Line SMMC-7721 in Vitro
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摘要: 目的 制备秦皮乙素并探讨其对人肝癌细胞株SMMC-7721增殖的影响。方法 采用聚酰胺柱层析法分离纯化秦皮乙素，采用薄层色谱法进行定性鉴别,高效液相色谱法进行纯度检测。采用MTT比色法观察秦皮乙素对人肝癌细胞株SMMC-7721增殖的影响。采用流式细胞仪分析秦皮乙素对SMMC-7721细胞周期及细胞凋亡的影响。结果 高效液相色谱法检测秦皮乙素的纯度为95%。MTT结果显示，IC50为2.24 mmol·L-1,并呈现时间-剂量依赖关系。流式细胞仪(FCM)检测表明, G2/M期细胞的比例减少，S期细胞的比例增加，与对照组相比差异显著(P<0.05)。结论 秦皮乙素可抑制人肝癌细胞株SMMC-7721的增殖，阻滞于S期，诱导肝癌细胞凋亡。Abstract: OBJECTIVE To prepare Esculetin and study the effect of Esculetin on proliferation of SMMC-7721 cell. METHODS Esculetin was isolated by polyamide chromatography. Esculetin was identified by TLC. The content of esculetin was tested by HPLC. MTT assay was adopted to evaluate cell proliferation. Distribution of cell cycle and apoptotic cell death were analyzed by flow cytometry. RESULTS The purity of Esculetin was 95% which was tested by HPLC. MTT assay showed that the inhibitory concentration 50% (IC50) of Esculetin was 2.24 mmol·L-1. Esculetin strongly inhibited growth of SMMC-7721 cell with dose-time dependent relationship. The result of FCM showed that G2/M phrase decreased and S phrase increased in SMMC-7721 cells with significant difference (P<0.05). CONCLUSION Esculetin could inhibit SMMC-7721 cell proliferation, disturbe the cell cycle progression and induce apoptosis.
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