论文部分内容阅读
目的观察BPA与B[a]P联合作用对3种乳腺上皮细胞MDM2及TP53基因表达的影响。方法采用细胞增殖荧光法检测环境相关剂量BPA对3种乳腺上皮细胞S期细胞比例的影响,采用实时荧光定量PCR和蛋白免疫印迹的方法分别检测环境相关剂量水平(10-9、10-7mol/L)BPA及其与10-6mol/L B[a]P联合作用对3种乳腺上皮细胞中MDM2和TP53基因及其编码的蛋白表达水平的影响。结果环境相关剂量的BPA可以引起MCF-7细胞S期细胞比例增高;环境相关剂量的BPA单独作用对3种类型乳腺上皮细胞MDM2和TP53基因表达水平无显著影响;B[a]P单独作用可以引起MCF-10A细胞中MDM2基因表达水平的轻微增加及MCF-7细胞中MDM2基因表达的显著增加;在MCF-7细胞中BPA与B[a]P联合作用可明显增加B[a]P所引起的MDM2基因表达上调,而在HMEC和MCF-10A细胞未发现该现象,蛋白水平与mRNA水平的改变基本一致,同时p53/mdm2比值降低。结论环境相关剂量的BPA与B[a]P联合作用可以在雌激素受体表达阳性的MCF-7细胞中引起MDM2基因及其蛋白产物表达水平的显著上调,提示环境暴露剂量的BPA可能通过雌激素受体依赖的作用途径增加化学致癌物致乳腺癌发生风险。
Objective To observe the effect of BPA combined with B [a] P on the expression of MDM2 and TP53 in three kinds of breast epithelial cells. Methods Cell proliferation fluorescence was used to detect the effect of environmental BPA on the proportion of S phase cells in three mammary epithelial cells. The levels of environmental related dose (10-9, 10-7 mol / L) were detected by real-time fluorescence quantitative PCR and Western blotting. L) BPA and their combination with 10-6mol / L [a] P on the expression of MDM2 and TP53 genes and their protein expression in three kinds of breast epithelial cells. Results Environmental-related doses of BPA caused an increase in the proportion of S-phase cells in MCF-7 cells. BPA alone in environmental-related doses had no significant effect on the expression of MDM2 and TP53 in three types of breast epithelial cells. B [a] P alone could Induced a slight increase of MDM2 gene expression in MCF-10A cells and a significant increase of MDM2 gene expression in MCF-7 cells; BPA combined with B [a] P significantly increased B [a] P in MCF-7 cells The up-regulation of MDM2 gene expression was not observed in HMEC and MCF-10A cells. The changes of protein level and mRNA level were basically the same, and the ratio of p53 / mdm2 was decreased. Conclusion The combination of BPA and B [a] P in environment-related dose can induce a significant up-regulation of MDM2 gene and its protein expression in estrogen receptor-positive MCF-7 cells, suggesting that the environmental exposure dose of BPA may be induced by estrogen Hormone receptor-dependent mechanisms of action increase the risk of chemical carcinogens causing breast cancer.