核因子-κB、p53及Bcl-2在铅诱导PC12细胞凋亡中的作用

目的探讨核因子κB(NFκ-B),p53及Bcl-2在铅诱导PC12细胞凋亡中的作用。方法应用细胞培养、流式细胞术、RT-PCR、Western Blot的方法分析NFκ-B、p53、Bcl-2基因的表达情况,研究醋酸铅诱导PC12细胞凋亡及其分子机理。结果PC12细胞经不同浓度醋酸铅(100、200和400μmol/L)处理24h后,用流式细胞仪检测其凋亡率(n=3),铅组凋亡率明显高于对照组,改变呈剂量依赖性。差异有显著性(P<0.05)。RT-PCR结果显示NFκ-B p65 mRNA、p53 mRNA转录水平随铅剂量升高而升高(P<0.05),两基因表达的改变呈剂量依赖性。Western Blot结果显示,表明随着铅浓度的增加,Bcl-2蛋白表达逐渐降低。结论铅诱导PC12细胞凋亡的这种效应可能与增强NFκ-B及p53基因的活性、降低Bcl-2基因的活性有关。

Study on the roles of nuclear factor-κB,p53 and Bcl-2 gene in lead acetate induced apoptosis in PC12 cells

Objective To investigate the roles of nuclear factor-κB(NF-κB),p53 and Bcl-2 gene in lead acetate induced apoptosis in PC12 cells.Methods Cell culture,Flow cytometry,RT-PCR and Western Blot were employed to analyze the expression of NF-κB, p53 and Bcl-2 gene,and clarify the effect of lead acetate on apoptosis in PC12 cells and related molecular mechanisms.Results The PC12 cells were exposed to lead acetate at varied concentrations(100μmol/L,200μmol/L and 400μmol/L) for 24 hours,Flow cytometry detected that the apoptosis rate of groups treated with lead acetate are higher than that of blank control groups with a dose-effect relationship(P<0.05).RT-PCR results show that the mRNA expression level of NF-κB p65,p53 increased accordingly.Western Blot results show that the protein expression of Bcl-2 gene decreased.Conclusion lead acetate induces apoptosis in PC12 cells,which may be related to activation of NF-κB and p53 gene and depression of Bcl-2 gene.