5-氮杂-2'-脱氧胞苷对胶质瘤细胞系U251凋亡的调节

5-Aza-2'-deoxycytidine regulates the apoptosis of glioma cell line U251

目的甲基化与胶质瘤的发生和发展关系密切,本研究的目的是探讨去甲基化试剂5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycytidine)对胶质瘤细胞系凋亡的调节。方法将浓度分别为10μM、25μM和50μM的5-氮杂-2'-脱氧胞苷加入胶质瘤细胞系U251细胞中,应用RT-PCR法检测U251细胞系经不同浓度的5-氮杂-2'-脱氧胞苷处理后细胞内胱氨酸蛋白酶-3(caspase-3)的表达,应用流式细胞术(AnnexinV-FITC染色)检测不同药物浓度处理组中U251细胞的凋亡率。结果 5-氮杂-2'-脱氧胞苷处理后,U251细胞内caspase-3的转录水平明显上调(P<0.01),具有较为明显的剂量依赖性。流式细胞术结果显示U251细胞经5-氮杂-2'-脱氧胞苷处理后细胞的凋亡率显著上调,且具有剂量依赖性。结论 5-氮杂-2'-脱氧胞苷处理具有剂量依赖性上调胶质瘤细胞系U251caspase-3表达,促进细胞凋亡。

Objective Many studies have shown that DNA methylation closely correlated with the occurrence and development of glioma, the purpose of this study is to investigate the effect of demethylating agent 5-Aza-2＇-deoxycytidine on the apoptosis of glioma cell line U251. Methods Different concentrations of 5-Aza-2＇-deoxyeytidine （10 μM, 25 μM and 50 μM） were used to treat glioma cell line U251, respectively. RT-PCR was used to detect the expression of easpase-3 mRNA in U251 ceils after 5-Aza-2＇-deoxycytidine treatment, and cell apoptosis rate was determined by flow cytometry using Annexin V-FITC method. Results 5-Aza-2＇-deoxycytidine treatment up-regulated the transcriptional level of caspase-3 significantly with significant dose-dependent （ P 〈0.01）, flow cytometry showed thai the apoptosis rate of U251 cells was up-regulated significantly after 5-Aza-2＇-deoxyeytidine treatment with a dose-dependent manner in glioma cell line U251. Conclusion 5-Aza-2＇-deoxycytidine treatment could up-regulate caspase-3 expression and promote the apoptosis rate of glioma cell line U251 with a dose-dependent manner.