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目的探讨去甲基化药物5-氮杂-2′脱氧胞苷(DAC)对结肠癌细胞及其表皮生长因子和卵泡抑素结构域跨膜蛋白(TMEFF2)基因表达的影响。方法以不同浓度的DAC药物处理结肠癌细胞Caco-2,采用Caspase 3/7试剂盒检测细胞凋亡率,同时提取药物处理前后的细胞总RNA,利用实时荧光定量RT-PCR的方法检测TMEFF2基因表达情况。结果DAC药物处理后Caco-2细胞凋亡率明显增加(P<0.05),且呈时间、药物浓度依赖性。去甲基化药物处理后,Caco-2细胞中TMEFF2基因m RNA表达水平明显升高(P<0.05)。结论 TMEFF2基因处于甲基化状态时,其在Caco-2细胞株中表达水平升高,提示TMEFF2基因启动子区的甲基化对下游m RNA的表达可能具有调控作用。
Objective To investigate the effects of 5-aza-2′-deoxycytidine (DAC), a demethylation drug, on the expression of colon cancer cells and its epidermal growth factor and follistatin domain transmembrane protein (TMEFF2) genes. Methods The colon cancer cells Caco-2 were treated with different concentrations of DAC drugs. The apoptosis rate was detected by Caspase 3/7 kit. Total RNA was extracted before and after drug treatment. The TMEFF2 gene was detected by real-time fluorescence quantitative RT-PCR. Expression of situation. Results The apoptosis rate of Caco-2 cells was significantly increased after DAC treatment (P<0.05), and it was time- and drug-dependent. After demethylation treatment, the expression of TMEFF2 mRNA in Caco-2 cells was significantly increased (P<0.05). Conclusion When TMEFF2 gene is in the methylated state, its expression level in Caco-2 cell line is increased, suggesting that the methylation of TMEFF2 gene promoter region may regulate the downstream m RNA expression.