5-Aza-CdR对胃癌细胞系BGC823生长及DAPK1基因甲基化的影响

Effects of 5-Aza-CdR on proliferation of human gastric cancer cell lines and abnormal methylation of DAPK1 gene

目的观察5-氮杂-2-脱氧胞苷(5-Aza-CdR)对体外培养的胃癌BGC823细胞增生和凋亡的影响及其对此细胞中DAPK1基因的甲基化状态的影响。方法选择浓度为1×10-6的5-Aza-CdR处理体外培养的BGC823细胞后,用Annexin-v-FITC凋亡检测药物处理后细胞凋亡情况;MSP法检测用药前后细胞中Apaf-1基因的甲基化状态;RT-PCR法检测用药前后细胞中DAPK1的mRNA表达的变化。结果在药物作用24、48、72 h后细胞的凋亡率分别为5.83±0.53、9.23±0.58、15.34±0.90,较对照组明显增加(P<0.05)。DAPK1基因的甲基化状态得到了逆转,DAPK-1基因的mRNA表达得到增加。结论5-Aza-CdR对BGC823细胞具有增生抑制作用,并促进细胞凋亡。DAPK1基因表达情况与其甲基化状态的改变有关。

[ Objective ] To observe the effects of 5-Aza-CdR on proliferation of human gastric cancer cell lines BGC823, methyla＇tion and epression of DAPK-1 gene. [ Methods] BGC823 cells were cultured in RPMI1640 and was treated with 5-Aza-CdR（ 1 ×10^ -6mol/L）. The proliferation of the cells was detected by flow cytometry. The methylation of DAPK1 in the two cell lines was detected by MSP, and the expression of DAPK1 was detected by RT-PCR. [Results] After BGC823 cells were exposed to 5-Aza-CdR, the growth of BGC823 cells was inhibited. The methylation and the loss of DAPK1 mRNA BGC823 cells were changed with 5-Aza-CdR. [ Conclusion ] 5-Aza-CdR can inhibited the growth of BGC823 cells and induced apoptosis, and changing the methylation of DAPK1 gene and improving DAPK1 ＇ s expression. The low expression of DAPK1 in BGC823 cells is because of methylation.