Objective To investigate the effect of 5-Aza-2'-deoxycitydine (5-Aza-dC) and (or) trichostatin A (TSA) on the methylation and mRNA expression of DLC-1 gene in gastric cancer cell line SNU-1.Methods Methylation-specific-polymerase chain reaction (MSP) was used to detect the methylation of DLC-1 gene in the gastric cancer cell lines and quantitative real-time polymerase chain reaction (q-PCR) was used to detect the expression of DLC-1 gene.Results In the control group,the SNU-1 cell line showed a characteristic DNA methylation status in promoter region of DLC-1 gene.After the treatments of different drugs for 72 hours,it was converted from methylation to unmethylation status in the 5-Aza-dC group.Meanwhile,the 5-Aza-dC + TSA combination group was similar to the 5-Aza-dC group;TSA group had no obvious influence on the methylation.The effect of different drugs on the expression of DLC-1 mRNA in gastric cancer cell lines SNU-1 showed that,in the control group,the expression of DLC-1 mRNA was low.Compared with a single drug (5-Aza-dc or TSA alone),the effect was significantly elevated after treatment with the combination of 5-Aza-dC + TSA (P ＜ 0.05).Conclusion 5-Aza-dC can reverse methylation status of DLC-1 gene promoter CpG island in the gastric cancer cell line SNU-1,and upregulate the expression of DLC-1 gene with TSA.%目的 探讨5-氮杂-2-脱氧胞苷(5-Aza-2'-deoxycitydine,5-Aza-dC)联合曲古抑菌素A(trichostatin A,TSA)对胃癌细胞株SNU-1中DLC-1基因的甲基化水平和mRNA表达的影响.方法 应用甲基化特异性聚合酶链式反应(MSP)和实时荧光定量PCR,检测5-Aza-dC单药或联合TSA作用于SNU-1细胞后,DLC-1基因的甲基化水平及DLC-1基因mRNA表达量.结果 对照组中DLC-1基因呈甲基化状态;5-Aza-dC药物干预组DLC-1基因出现明显的非甲基化条带,甲基化条带亮度减弱;TSA组条带无明显变化;5-Aza-dC+ TSA组DLC-1基因甲基化条带明显减弱,且出现非甲基化条带.5-Aza-dC组DLC-1 mRNA表达量是对照组的2.22倍,TSA组DLC-1 mRNA表达量是对照组的2.20倍,5-Aza-dC+ TSA组DLC-1 mRNA表达量是对照组的4.50倍(P＜0.05).结论 5-Aza-dC能诱导胃癌细胞系SNU-1中DLC-1基因去甲基化,与TSA联用可提高DLC-1 mRNA表达.
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