清毒栓提取物通过调节Zeste基因增强子同源物2抑制SiHa细胞活性的研究  

Study on Qingdushuan extract inhibiting the activity of SiHa cells by regulating EZH2

作　　者：李美 楼姣英[2] 耿韦华 乔晓叶 LI Mei;LOU Jiaoying;GENG Weihua;QIAO Xiaoye(Beijing University of Chinese Medicine,100029,Beijing,China)

机构地区：[1]北京中医药大学,100029 [2]北京中医药大学东方医院妇科

出　　处：《环球中医药》2021年第4期579-586,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81473515)。

摘　　要：目的研究清毒栓提取物对人子宫颈鳞癌(SiHa)细胞活性的影响,并基于Zeste基因增强子同源物2(enhancer of zeste homolog 2,EZH2)探讨其可能的作用机制。方法取稳定并进入对数生长期的SiHa细胞,采用血清药理学方法制备的清毒栓含药血清和EZH2抑制剂GSK126进行干预,使用八肽胆囊收缩素、实时荧光定量PCR法、蛋白免疫印迹法、染色质免疫沉淀技术等技术,研究清毒栓含药血清对SiHa细胞活性、增殖与凋亡、叉状头/翅膀状螺旋转录因子(Forkhead box P3,Foxp3)表达与其组蛋白甲基化水平、β-连环蛋白通路、EZH2表达影响。结果清毒栓含药血清促进Foxp3启动子区组蛋白甲基化水平,并可增加EZH2含量;而Foxp3过表达质粒和EZH2抑制剂GSK126可以逆转清毒栓含药血清诱导的Foxp3表达下调,增殖凋亡相关蛋白表达改变以及SiHa细胞活性下调。结论清毒栓对SiHa细胞的抑制作用是通过增加EZH2含量促进Foxp3启动子组蛋白甲基化来改变Foxp3表达,从而抑制SiHa细胞中β-连环蛋白通路实现的。Objective To study the effect of Qingdushuan extract on the activity of SiHa cells in cervical cancer.And to explore its possible mechanism based on EZH2(Zeste gene enhancer homolog2).Methods The SiHa cells that were stable and entered the logarithmic growth stage were taken and the Qingdushuan drug-containing serum prepared by serum pharmacology method and the EZH2 inhibitor GSK126 were used for intervention.The CCK-8,qRT-PCR,WB,CHIP and other techniques were used to study the effects of Qingdushuan drug-containing serum on the activity of SiHa cells,proliferation and apoptosis,Foxp3 expression and their histone methylation levels,β-catenin pathway,and EZH2 expression.Results Qingdushuan drug-containing serum could promoted histone methylation level of Foxp3 promoter region and increased the content of EZH2.While Foxp3 overexpressed plasmid and EZH2 inhibitor GSK126 could reverse the expression down-regulation of Foxp3,the expression changes of in apoptosis-associated proteins and the activity down-regulation of SiHa cells.Conclusion The inhibitory effect of Qingdushuan on SiHa cells was achieved by increasing the content of EZH2 and promoting the histone methylation of Foxp3 promoter to change the expression of Foxp3,thereby inhibitingβ-catenin pathway in SiHa cells.

关 键 词：清毒栓含药血清 宫颈癌 Zeste基因增强子同源物2 叉头状螺旋转录因子 β-连环蛋白通路 

分 类 号：R285.5[医药卫生—中药学]