摘要
目的研究绿茶提取物表没食子儿茶素-3-没食子酸酯(Epigallocatechin-3 gallste,EGCG)对人卵巢癌HO-8910细胞增殖及细胞内Wnt/β-catenin信号通路相关基因表达的影响,探讨EGCG抑制卵巢癌细胞生长的机制。方法用不同浓度的EGCG(10、20和40μg/ml)处理体外培养的人卵巢癌HO-8910细胞不同时间(24、48和72 h),采用MTT法检测细胞的增殖活力;流式细胞术检测细胞周期的变化;RT-PCR和Western blot分别检测细胞中β-catenin和下游靶基因CyclinD1 mRNA的转录水平和蛋白的表达水平。结果 EGCG可明显抑制HO-8910细胞的增殖活力,且抑制作用呈剂量-时间依赖性(P<0.05);40μg/ml EGCG干预后,HO-8910细胞主要阻滞于G0/G1期,且在48 h阻滞作用最为明显;EGCG可显著降低HO-8910细胞中β-catenin和CyclinD1基因mRNA的转录水平和蛋白的表达水平,且呈剂量-时间依赖性(P<0.01)。结论 EGCG可抑制HO-8910细胞的增殖,其机制可能与其抑制Wnt/β-catenin信号通路的活性有关,提示EGCG可能在卵巢癌的治疗中具有一定的应用前景。
Objective To observe the effect of epigallocatechin-3-gallste（EGCG）,an extract from green tea,on proliferation of human ovarian cancer HO-8910 cells as well as expression of Wnt / β-catenin signaling pathway-associated genes,and investigate the potential mechanism of EGCG in inhibiting the growth of ovarian cancer cells.Methods HO-8910 cells cultured in vitro were treated with EGCG at various concentrations（10,20 and 40 μg / ml） for various hours（24,48 and 72 h）,then determined for proliferative activity by MTT method,for cell cycle by flow cytometry,for transcription levels of β-catenin and CyclinD1 mRNAs by RT-PCR,and for expression levels of β-catenin and CyclinD1 by Western blot.Results EGCG showed significantly dose-and time-dependent inhibitory effect on the proliferative activity of HO-8910 cells（P 0.05）.G0 / G1 arrest was observed in HO-8910 cells treated with 40 μg / ml EGCG,of which the most obvious one appeared 48 h after treatment.EGCG decreased the transcription levels of β-catenin and CyclinD1 mRNAs and expression levels of β-catenin and CyclinD1 significantly,in dose-and time-dependent modes（P〈0.01）.Conclusion EGCG inhibited the proliferation of HO-8910 cells by a mechanism which might be associated with inhibiting the activity of Wnt / β-catenin signaling pathway,indicating a potential prospect of application of EGCG to the therapy of ovarian cancer.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第9期1165-1170,共6页
Chinese Journal of Biologicals
基金
高等学校博士学科点专项科研项目(200806310001)
