肾小管上皮细胞间质转分化过程中E-cadherin和α-SMA基因启动子区甲基化状态研究

Methylation status in E-cadherin and α-SMA gene promoter region during renal tubular epithelial-mesenchymal transition

目的:了解大鼠肾小管上皮细胞间质转分化与E-钙黏附蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA)基因启动子区甲基化状态之间的关系。方法:将大鼠肾小管上皮细胞分为对照组和TGF-β1处理组。对两组细胞采用免疫细胞化学方法检测E-cadherin和α-SMA蛋白表达情况,用ELISA法检测培养液中Ⅰ型胶原蛋白的含量。同时分别提取两组细胞中的基因组DNA,应用焦磷酸测序技术对E-cadherin和α-SMA基因启动子区甲基化水平进行检测。结果:TGF-β1处理组细胞中Ecadherin基因启动子区有2个CpG位点甲基化频率高于对照组,而α-SMA基因启动子区有2个CpG位点甲基化频率低于对照组。结论:E-cadherin和α-SMA基因启动子区甲基化状态的改变参与了肾小管上皮细胞间质转分化过程。

Objective：To investigate the relationship between renal tubular epithelial-mesenchymal transition and promoter methylation status of E-cadherin and α-SMA gene. Methods： Renal tubular epithelial cells of rat were divided into control group and TGF-β1 treated group. E-cadherin and α-SMA expression were detected by immunocytochemical staining; type Ⅰ collagen concentration in culture medium was tested by ELISA method;meanwhile genomic DNA was extracted, then the E-cadherin and α-SMA promoter methylation status was detected using pyrosequencing technique. Results： There was higher methylation level and lower methylation level in promoter region ofα-SMA gene in TGF-β1 group than in control group. Conclusion：Methylation changes in E-cadherin andα-SMA gene promoter region is involved in renal tubular epithelial-mesenchymal transition.