5-氮杂-2'-脱氧胞苷对胃癌AGS细胞增殖、凋亡的影响及机制探讨  

Inhibition effect and mechanism of 5-Aza-CdR on proliferation and apoptosis in gastric cancer cell line AGS

作　　者：袁昌劲[1] 唐求[1] 王辉[1] 陈锐[2] 

机构地区：[1]武汉市中心医院,武汉430014 [2]广州中医药大学第一附属医院

出　　处：《山东医药》2011年第14期3-5,共3页Shandong Medical Journal

摘　　要：目的观察5-氮杂-2'-脱氧胞苷(5-Aza-CdR)对胃癌AGS细胞增殖、凋亡的影响,并探讨其机制。方法分别用低、中、高浓度5-Aza-CdR处理AGS,对照组不处理。CCK-8检测AGS细胞增殖状态,流式细胞仪检测AGS细胞周期及凋亡率,MSP检测AGS中DAPK基因启动子甲基化状态,RT-PCR和Western blot法分别检测AGS中的DAPKmRNA及其蛋白。结果低、中、高浓度组AGS细胞增殖速度显著低于对照组(P均<0.05),并呈浓度、时间依赖性(P均<0.05)。5-Aza-CdR使AGS细胞停滞在G0/G1期(P均<0.05),细胞凋亡率增加(P均<0.05)。对照组DAPK基因启动子表现甲基化状态,低、中、高浓度组DAPK基因启动子被逆转成未甲基化状态。低、中、高浓度组AGS细胞DAPK mRNA及蛋白表达量均显著高于对照组(P均<0.05),并呈浓度依赖性。结论 5-Aza-CdR可抑制AGS细胞增殖、促进其凋亡,可能与逆转DAPK基因启动子甲基化状态并使其重新表达有关。Objective To study the inhibition effect of 5-Aza-2′deoxycytidine（5-Aza-CdR） on proliferation and apoptosis of gastric cancer cell line AGS and explore its possible mechanisms.Methods Low,median and high concentrations of 5-Aza-CdR were used to treat gastric cancer cell line AGS.Cell counting kit-8 was employed to detect the proliferation of gastric cancer cell line AGS;flow cytometer was used to detect the cell cycle and apoptosis;MSP,RT-PCR and Western Blot were applied to detect the methylation status and gene expression of DAPK pre-and post-treatment respectively.Results The proliferation rates in low,median and high concentration groups were significantly lower than control group in a dose-and time-dependent manner（P all 0.05）.AGS cells stagnated in G0/G1 after 5-Aza-CdR treatment with promoted apoptosis rates（P all 0.05）.DAPK gene＇ s promoter was methylated,while it was reversed unmethylated in low,median and high concentration groups.The gene expression of DAPK in low,median and high concentration groups were significantly higher than control group in a concentration-dependent manner（P all 0.05）.Conclusion 5-Aza-CdR can inhibit the proliferation and promote the apoptosis of gastric cancer line AGS by reversing the promoter＇ s methylation status and gene expression of DAPK gene.

关 键 词：5-氮杂-2'-脱氧胞苷 死亡相关蛋白激酶 甲基化状态 胃癌 胃癌细胞 

分 类 号：R735.2[医药卫生—肿瘤]