小檗碱对人卵巢癌细胞SKOV3 增殖及凋亡机制的研究

目的:探讨小檗碱对人卵巢癌细胞(SKOV3)增殖及凋亡的影响。方法:MTT 法检测细胞增殖;流式细胞仪Annexin V/ PI 双染色法和透射电子显微镜检测细胞凋亡情况;甲基化特异性PCR 分析hMLH1 基因启动子区CpG 岛的甲基化状态;实时荧光定量RT-PCR 检测Bcl-2、Bax、Survivin 和hMLH1 mRNA 基因的表达。结果:小檗碱对卵巢癌SKOV3 细胞增殖有明显的抑制作用(P<0.05),呈剂量和时间依赖性。当与顺铂联用时,小檗碱对卵巢癌细胞有协同抗癌作用。小檗碱可明显诱导SKOV3 细胞凋亡,并下调Bcl-2、Survivin 基因及上调Bax 基因的表达。此外,小檗碱能恢复hMLH1 启动子的甲基化状态及增强hMLH1 mRNA 的表达。结论:小檗碱可抑制卵巢癌细胞增殖及诱导细胞凋亡,小檗碱可协同增强抗癌药物顺铂的抗肿瘤作用。

Research of berberine to proliferation and apoptotic mechanism of human ovarian cancer cell line SKOV3

Objective:To investigate the effect of berberine on the proliferation and apoptosis of human ovarian cancer cell(SKOV3). Methods:Cell proliferation was detected by MTT method.The cell apoptosis was detected by FCM Annexin V/ PI double staining and transmission electron microscopy.The methylation status of hMLH1 gene promoter CpG island was analyzed by methylation specific PCR.The expression of Bcl-2, Bax, Survivin and hMLH1 gene mRNA were detected by real-time fluorescent quantitative RT-PCR.Results:The berberine could significantly inhibit the proliferation of ovarian cancer SKOV3 cells(P<0.05) in dose-and time-de-pendent manner.When combined with cisplatin, berberine showed synergistic anticancer effects.Berberine could induce SKOV3 cells apoptosis significantly, it might lower the expression of Bcl-2 and Survivin gene and enhance the expression of Bax gene.In addition,berberine could restore the hMLH1 promoter methylation status and increase the expression of hMLH1 mRNA.Conclusion:Berberine can inhibit the proliferation of ovarian cancer cells and induce apoptosis, which show that the synergistic enhancement anticancer effects with cisplatin.