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[目的]研究去甲基化5-氮杂-2′-脱氧胞苷(5-Aza-CdR)对人结肠腺癌细胞株Caco-2细胞生长周期及凋亡的影响,探讨其临床治疗的可能性。[方法]分别使用0.4、1.6、6.4、25.6、102.4μmol/L浓度的5-Aza-CdR处理Caco-2细胞;通过MTT检测5-Aza-CdR对Caco-2细胞存活率的影响;应用流式细胞检测5-Aza-CdR对Caco-2细胞生长周期及凋亡的影响;RT-PCR检测处理前后抑癌基因RASSF1A mRNA表达的改变。[结果]1.6μmol/L浓度的5-Aza-CdR可以明显的抑制Caco-2细胞的增殖,细胞周期中处于G0/G1期的细胞明显的增多,阻滞于G1期,凋亡率增高;5-Aza-CdR的作用与其浓度、时间在一定范围内呈正相关。5-Aza-CdR处理后,无RASSF1A表达的Caco-2细胞可检测出基因RASSF1A的重新表达。[结论]5-Aza-CdR可消除某些抑癌基因启动子甲基化状态,使其重新表达而抑制Caco-2细胞的生长,并促进其凋亡。
[Objective] To investigate the effect of 5-Aza-2’-deoxycytidine (5-Aza-CdR) on the growth cycle and apoptosis of human colon adenocarcinoma cell line Caco-2 and to investigate its clinical treatment possibility. [Methods] Caco-2 cells were treated with 5-Aza-CdR at the concentrations of 0.4,1.6,6.4,25.6,102.4μmol / L, respectively. The effect of 5-Aza-CdR on the viability of Caco- The effect of 5-Aza-CdR on the growth cycle and apoptosis of Caco-2 cells was detected by RT-PCR and the expression of the tumor suppressor gene RASSF1A mRNA was detected by RT-PCR. [Result] The concentration of 1.6μmol / L 5-Aza-CdR significantly inhibited the proliferation of Caco-2 cells. The number of cells in G0 / G1 phase in the cell cycle increased significantly, arrested in G1 phase and the apoptosis rate increased. The role of 5-Aza-CdR was positively correlated with its concentration and time within a certain range. After 5-Aza-CdR treatment, re-expression of gene RASSF1A was detected in Caco-2 cells without RASSF1A expression. [Conclusion] 5-Aza-CdR can eliminate the methylation status of some tumor suppressor gene promoters, re-express them and inhibit the growth of Caco-2 cells and promote their apoptosis.