表没食子儿茶素没食子酸酯促进急性髓系白血病细胞凋亡的机制  

Epigallocatechin gallate induces CHD5 gene demethylation to promote acute myeloid leukemia cell apoptosis in vitro by regulating p19^Arf-p53-p21^Cip1 signaling pathway

作　　者：吴明彩[1,2] 蒋明 薛梦雅 李青 程彬[1] 黄梦珠 徐蕾 章尧[1,2] WU Mingcai;JIANG Ming;XUE Mengya;LI Qing;CHENG Bing;HUANG Mengzhu;XU Lei;ZHANG Yao(Department of Biochemistry and Molecular Biology,Wannan Medical College,Wuhu 241002,China;Anhui Provincial Key Laboratory of Active Biological Macromolecules,Wuhu 241002,China;Wuhu Second Sanatorium for Retired Cadres,Anhui Provincial Military Command,Wuhu 241002,China)

机构地区：[1]皖南医学院生物化学与分子生物学教研室,安徽芜湖241002 [2]安徽省活性大分子重点实验室,安徽芜湖241002 [3]安徽省军区芜湖市第二离职干部休养所,安徽芜湖241002

出　　处：《南方医科大学学报》2020年第9期1230-1238,共9页Journal of Southern Medical University

基　　金：国家自然科学基金(31971199);安徽高校自然科学研究项目重点项目(KJ2018A0262);安徽省高等学校省级质量工程教学研究重点项目(2019jyxm025);皖南医学院博士科研启动基金项目(WYRCQD2018001);皖南医学院青年骨干人才项目(wyqnyx201906);安徽省大学生创新训练项目(S201910368088,201810368063,201810368064)。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)诱导CHD5基因去甲基化促进KG-1、THP-1细胞凋亡的相关机制。方法设实验组及正常对照组,实验组:不同浓度(25、50、75、100、150μg/mL)EGCG处理作用于KG-1、THP-1细胞,正常对照组:不加EGCG处理。MSP法检测KG-1、THP-1细胞CHD5基因甲基化;MTT法检测作用48 h后细胞增殖;流式细胞术检测作用48 h后细胞周期和细胞凋亡状况;RT-qPCR、Western blot检测DNMT1、CHD5、p19^Arf、p53、p21^Cip1基因和蛋白表达。结果EGCG呈现剂量依赖性逆转了KG-1、THP-1细胞CHD5基因高甲基化;EGCG以剂量依赖性的方式抑制KG-1、THP-1细胞增殖(P<0.05);EGCG诱导细胞周期停滞于G1期,促进细胞凋亡;EGCG以剂量依赖性的方式下调DNMT1的mRNA和蛋白表达,上调CHD5、p19^Arf、p53、p21^Cip1的mRNA和蛋白表达(P<0.05)。结论EGCG可通过下调DNMT1降低KG-1、THP-1细胞CHD5基因高甲基化,恢复CHD5基因表达,从而上调p19^Arf、p53、p21^Cip1表达诱发细胞凋亡。Objective To investigate the mechanism by which epigallocatechin gallate(EGCG)induces CHD5 gene demethylation and promotes the apoptosis of acute myeloid leukemia KG-1 and THP-1 cell lines.Methods KG-1 and THP-1 cells treated with 25,50,75,100 or 150μg/mL EGCG for 48 h were examined for CHD5 gene methylation using MSP and for cell proliferation using MTT assay.The changes in cell cycle and apoptosis of the two cell lines after treatment with EGCG for 48 h were detected using flow cytometry.The mRNA and protein expressions of DNMT1,CHD5,p19^Arf,p53 and p21^Cip1 in the cells were detected using RT-quantitative PCR and Western blot.Results EGCG dose-dependently reversed hypermethylation of CHD5 gene and reduced the cell viability in both KG-1 and THP-1 cells(P<0.05).EGCG treatment caused obvious cell cycle arrest in G1 phase,significantly increased cell apoptosis,downregulated the expression of DNMT1 and upregulated the expressions of CHD5,p19^Arf,p53 and p21^Cip1 in KG-1 and THP-1 cells(P<0.05).Conclusion EGCG reduces hypermethylation of CHD5 gene in KG-1 and THP-1 cells by downregulating DNMT1 to restore its expression,which results in upregulated expressions of p19^Arf,p53 and p21^Cip1 and induces cell apoptosis.

关 键 词：表没食子儿茶素没食子酸 CHD5基因 甲基化 p19^Arf-p53-p21^Cip1信号通路 急性髓系白血病 

分 类 号：R285[医药卫生—中药学]