5-Aza-CdR对MDA-MB-231细胞PPAR-γ基因甲基化的影响  

Effect of 5-Aza-2'-deoxycytidine on methylation of PPAR-γ gene in MDA-MB-231 cell

作　　者：董家书[1] 韦美德[1] 周格琛[1] 邓开凤[1] 戴盛明[1] DONG Jiashu;WEI Meide;ZHOU Gechen;DENG Kaifeng;DAI Shengming(Medical Science Eaboratory,the Fourth Affiliated Hospital of Guangxi Medical University,Liuzhou Guangxi China,54500)

机构地区：[1]广西医科大学附属第四医院检验科,广西柳州545005

出　　处：《分子诊断与治疗杂志》2018年第4期252-256,共5页Journal of Molecular Diagnostics and Therapy

基　　金：BEAMing技术定量检测乳腺癌患者外周血PPAR-γ甲基化及临床应用(2015J030510)

摘　　要：目的研究去甲基化药物5-氮杂-2-脱氧胞嘧啶(5-aza-2.-deoxycytidine,5-Aza-CdR)对三阴性乳腺癌(triple-negative breast cancer,TNBC)细胞MDA-MB-231凋亡及过氧化物酶体增殖物激活受体-γ(peroxisome proliferator-activated receptor-γ,PPAR-γ)基因表达的影响。方法 5-Aza-CdR作用MDA-MB-231细胞后,流式细胞术检测MDA-MB-231细胞凋亡,实时荧光定量PCR(real-time fluorescent quantive PCR,q PCR)检测PPAR-γ m RNA的表达,甲基特异性PCR(methylmion specific PCR,MSP)检测PPAR-γ甲基化的改变。结果流式细胞结果表明,随着药物浓度增加,细胞处理48 h后,凋亡率增加,呈剂量依赖性。5、10、15和20μmol/L组凋亡率分别为(14.1±2.3)%、(25.4±3.3)%、(32.7±2.8)%、(43.1±1.9)%,与对照组的(6.9±0.8)%相比差异有统计学意义(P<0.05);q PCR结果显示,随着药物浓度增加,PPAR-γ m RNA表达上调;MSP显示PPAR-γ甲基化水平降低。结论 5-Aza-CdR能降低PPAR-γ甲基化状态,使PPAR-γ表达增加,促进MDA-MB-231细胞凋亡。PPAR-γ甲基化水平改变可能是引起MDA-MB-231细胞生物学改变的机制之一。Objective To investigate the effects of 5-aza-2＇-deoxycytidine（5-Aza-CdR）on the proliferation of MDA-MB-231 cells and methylation of peroxisome proliferator-activated receptor-γ（PPAR-γ）.Methods The expression and methylation status of PPAR-γ m RNA was detected by real-time fluorescent quantitive PCR（q PCR）,the apoptosis of MDA-MB-231 cells was examined by flow cytometry（FCM）.Results Flow cytometry results showed that with the increase of drug concentration,the apoptosis rate increased after 48 h of cell treatment in a dose-dependent manner. The apoptosis rate was（14.1±2＇3）% in 5μmol/L 5-Aza-CdR group,（25.4±3.3）% in 10 μmol/L 5-Aza-CdR group,（32＇7±2＇8）% in 15 μmol/L 5-AzaCdR group and（43.1±1.9）% in 20 μmol/L 5-Aza-CdR group,significantly higher than that in the control group（6.9±0.8）%. The expression of PPAR-γ was increased and the methylation level of PPAR-γ was decreased by the treatment of different concentration of 5-Aza-CdR（5,10,15 and 20 μmol/L）. Conclusion 5-Aza-CdR can decrease the methylation level of PPAR-γ,up-regulate the expression of PPAR-γ and promote MDA-MB-231 cells apoptosis. The changes of methylation level of PPAR-γ may be one of the mechanisms that cause biological changes in MDA-MB-231 cells.

关 键 词：5-AZA-CDR MDA-MB-231细胞 PPAR-Γ 甲基化 

分 类 号：R737.9[医药卫生—肿瘤]