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目的研究香加皮中宝藿苷-Ⅰ对人食管癌细胞Eca-109细胞周期的影响及其作用机制。方法采用MTT法分析不同质量浓度宝藿苷-Ⅰ(12.5、25、50μg/mL)分别作用24、48、72h后,对Eca-109细胞增殖的影响;流式细胞术分析宝藿苷-Ⅰ对Eca-109细胞周期的影响;RT-PCR技术检测宝藿苷-Ⅰ对Eca-109细胞Cyclin B1mRNA表达的影响;Western blotting方法检测宝藿苷-Ⅰ对Eca-109细胞Cyclin B1蛋白表达的影响。结果25、50μg/mL宝藿苷-Ⅰ均可明显抑制Eca-109细胞的增殖(P<0.05),作用48h的IC50为24.8μg/mL。25、50μg/mL宝藿苷-Ⅰ作用48h后,随宝藿苷-Ⅰ质量浓度的增加,Eca-109细胞的G0/G1期细胞比例明显下降,G2/M期细胞比例明显提高,细胞的Cyclin B1蛋白及mRNA表达水平降低,与对照组相比均有显著性差异(P<0.01)。结论香加皮宝藿苷-Ⅰ可抑制Eca-109细胞增殖,使细胞周期阻滞,该作用可能与下调细胞Cyclin B1表达有关。
Objective To study the effect of icariin-Ⅰ on the cell cycle of human esophageal cancer cell line Eca-109 and its mechanism. Methods MTT assay was used to analyze the effects of different doses of icoxabine-Ⅰ (12.5,25 and 50 μg / mL) on the proliferation of Eca-109 cells after 24h, 48h and 72h, respectively. The results of flow cytometry On the cell cycle of Eca-109 cells; the effect of treasure-glycoside-I on the expression of Cyclin B1 mRNA in Eca-109 cells was detected by RT-PCR; the effect of Po-glycoside-I on the expression of Cyclin B1 in Eca-109 cells was detected by Western blotting . Results 25,50μg / mL of treasure glycoside-Ⅰ could significantly inhibit the proliferation of Eca-109 cells (P <0.05), the IC50 of 48h was 24.8μg / mL. After 48 and 50μg / mL of glucoraphanin-Ⅰ for 48h, the percentage of cells in G0 / G1 phase of Eca-109 cells decreased significantly and the proportion of cells in G2 / M phase increased obviously Cyclin B1 protein and mRNA expression levels decreased, compared with the control group were significantly different (P <0.01). Conclusions Comboglycidin-Ⅰ can inhibit the proliferation of Eca-109 cells and block the cell cycle, which may be related to the down-regulation of Cyclin B1 expression.