去甲基化药物DAC对结肠癌细胞及其TMEFF2基因表达的影响

Effects of DAC on the expression of TMEFF2 gene and the state of colon cancer

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 explore the effects of 5-aza-2′-deoxycytidine（DAC） on the state and expression of transmembrane protein with EGF-like and two follistatin-like domains 2（TMEFF2） in colon cancer cell. Methods Caco-2 cells was treated with demethylation agent DAC,and then measured the cell death rates by caspase 3 / 7 detection kit,and determined the expression changes of TMEFF2 using Real-time RT-PCR. Results The treatment with DAC led to enhance the cell apoptosis and could potentially inhibit the Caco-2 cell growth in dose-dependent and time-dependent manners. The Caco-2 cells treated with DAC could have increased TMEFF2 m RNA expression level as compared with control. Conclusion Methylation in the promoter region of some of the cancer-related genes could suppress the m RNA expression and release the tumor cells from apoptosis or promote the tumor initiation and progression.