普鲁卡因抑制人结肠癌细胞增殖与DNA甲基化

Procaine inhibits the proliferation and DNA methylation in human colon cancer cells

目的分析普鲁卡因(PCA)对Septin9甲基化状态和表达水平的影响,探讨PCA抑制结肠癌DLD-1、HCT116细胞增殖的分子机制。方法不同浓度PCA对结肠癌细胞处理不同时间,MTS法检测细胞增殖活性;流式细胞术、Transwell实验检测PCA对细胞周期、凋亡和迁移能力的影响;BGS法、qRT-PCR检测PCA作用前后Septin9甲基化状态和表达水平。结果PCA对结肠癌细胞增殖抑制呈浓度和时间依赖效应(P<0.05)。与对照组比较,PCA处理后将细胞生长阻滞在G_2/M期、显著诱导凋亡且抑制细胞的迁移(P<0.05)。PCA处理后Septin9甲基化程度下降,Septin9 mRNA表达量上升,差异均有统计学意义(P<0.05)。结论PCA可以通过逆转Septin9甲基化,抑制结肠癌细胞的增殖、迁移。

Objective To analyze the effect of procaine(PCA)on the methylation status and expression level of Septin9,and to explore the molecular mechanism of PCA inhibiting the proliferation of colon cancer DLD-1 and HCT116 cells.Methods Different concentrations of PCA treated colon cancer cells at different times,and MTS method was used to detect cell proliferation activity.Flow cytometry and Transwell experiments were used to detect the effects of PCA on cell cycle,apoptosis and migration ability;BGS method and qRT-PCR were used to detect the methylation status and expression level of Septin9 before and after PCA treatment.Results PCA inhibited the proliferation of DLD-1 and HCT116 cells in a concentration-and time-dependent manner(P<0.05).Compared with the control group,PCA treatment blocked cell growth in G2/M phase,significantly induced apoptosis and inhibited cell migration(P<0.05).After PCA treatment,the degree of Septin9 methylation decreased,the expression of Septin9 mRNA increased,and the differences were statistically significant(P<0.05).Conclusion PCA can inhibit the proliferation and migration of colon cancer cells by reversing Septin9 methylation.