8-Br-cAMP和槲皮素对Eca-109细胞iNOS基因拷贝及Caspase-3表达的影响

Expression of iNOS gene and Caspase-3 in Eca-109 cell apoptosis induced by 8-Br-cAMP and quercetin

目的 :观察 8 Br cAMP和槲皮素对Eca 10 9细胞iNOS基因拷贝及Caspase 3表达的影响。方法 :将贴壁培养的Eca 10 9细胞随机分成 3组 ,Br组 :加 8 Br cAMP至终浓度为 2× 10 5mol/L ;Q组 :加槲皮素至终浓度为 4 3μmol/L ;C组 :不加任何药物 ,只加入新培养液培养。 3组细胞同时培养 4 8h。采用原位斑点印迹法显示iNOS基因拷贝和mRNA的表达 ,采用细胞免疫化学技术显示Caspase 3的表达。结果 :Br和Q组较C组iNOS基因拷贝和mRNA表达信号较强。Br和Q组细胞内Caspase 3免疫反应 (IR)性较强 ,Br和Q组与对照组相比 ,存在显著性差异 (P 0 .0 1) ;Br组的Caspase 3 IR强于槲皮素组 (P <0 .0 1)。结论 :8 Br cAMP和槲皮素皆可上调iNOS基因的扩增和表达 ,最终可都通过上调Caspase 3的表达 ,诱导Eca 10 9细胞凋亡。

Aim:To study the amplification and expression of iNOS gene and its role in Eca 109 cell apoptosis.Methods: The cultured Eca 109 cells were randomly divided into 3 groups: 8 Br cAMP (Br)group, Quercetin (Q)group, and Control (C)group with no any drugs.The 3 group cells were cultured for 48 h under the same condition.The expression and copies of iNOS gene were demonstrated by DNA/RNA dot blot and the caspase 3 immunoractivity(IR) by immunohistochemistry.Results: Compared with the C group, the gene copies and mRNA dot blot signals and caspase 3 IR were stron ger in the Br and Q groups ( P <0.01); Caspase 3 IR was higher in Br group than that in Q group( P <0.01).Conclusion: Both 8 Br cAMP and quercetin could up regulate iNOS gene copies and expression, and finally induce Eca 109 cell apoptosis through up regulating expression of Caspase 3.