白术内酯Ⅰ抑制卵巢癌SK-OV-3与OVCAR-3细胞增殖作用机制的研究

摘要

Objective To investigate the mechanisms and proliferation inhibitory effects of atractylenolide Ⅰ on SK-OV-3 and OVCAR-3 ovarian cancer cell.Methods SK-OV-3 and OVCAR-3 cells were treated with atractylenolide I with various concentrations at 24 hours,48 hours and 72 hours,and the changes in proliferation were detected by MTT assay.The cell cycles were measured by PI staining and flow cytometry,and the expressions of cyclin D1 and CDK1 were detected by ELISA assay.Western blot was then applied to investigate the effects of atractylenolide Ⅰ on PI3K/AKT signaling pathway in SK-OV-3 and OVCAR-3 cells.Results Atractylenolide I could significantly inhibit the proliferation of SK-OV-3 and OVCAR-3 cells,and its inhibitory effects were concentration and time dependent.In addition,atractylenolide I could also significantly reduce the proportion of cells in S phase and increase the proportion of cells in G2/M phase,and these effects were associated with the down-regulation of CDK1.The results of Western blot indicated that PI3K/AKT signaling pathway was involved in the inhibitory effects of atractylenolide Ⅰ on proliferation and cell cycle.Conclusion Atractylenolide I can down-regulate the expression of CDK1 in ovarian cancer SK-OV-3 and OVCAR-3 cells through PI3K/AKT pathway,which led to cell cycle arrest in G2/M phase,and played an important role in proliferation inhibition of tumor cells.%目的 探讨白术内酯Ⅰ抑制卵巢癌SK-OV-3与OVCAR-3细胞增殖的作用及其分子机制.方法 MTT实验检测不同浓度白术内酯Ⅰ作用人卵巢癌SK-OV-3与OVCAR-3细胞24h、48 h、72 h后,白术内酯Ⅰ对卵巢癌细胞增殖的影响;流式细胞技术与PI染色检测白术内酯Ⅰ对卵巢癌SK-OV-3与OVCAR-3细胞周期的影响,ELISA实验检测白术内酯Ⅰ对卵巢癌SK-OV-3与OVCAR-3细胞cyclin D1与CDK1表达的影响;Western blot测定白术内酯Ⅰ对卵巢癌SK-OV-3与OVCAR-3细胞PI3 K/AKT信号通路的影响.结果 白术内酯Ⅰ可显著抑制SK-OV-3与OVCAR-3细胞增殖,且白术内酯Ⅰ对SK-OV-3与OVCAR-3细胞的增殖抑制作用具有浓度和时间依赖性;此外,白术内酯Ⅰ还可显著降低SK-OV-3与OVCAR-3细胞的S期细胞比例,增加G2/M期细胞比例,且白术内酯Ⅰ对SK-OV-3与OVCAR-3细胞周期的调控与其抑制CDK1的表达有关;Western blot实验进一步探讨了白术内酯Ⅰ抑制细胞增殖和调控细胞周期与PI3 K/AKT信号通路有关.结论 白术内酯Ⅰ可通过PI3 K/A KT途径下调卵巢癌SK-OV-3与OVCAR-3细胞CDK1的表达,从而使细胞阻滞于G2/M期,进而发挥肿瘤细胞增殖抑制作用.