Deming Li, Ziwei Li, Xueqiang Guo, Cunshuan Xu
Key Laboratory for Cell Differentiation Regulation, Xinxiang, China ; College of Life Science, Henan Normal University, Xinxiang, China.
Cell J. 2015 Fall;17(3):576-81. doi: 10.22074/cellj.2015.20. Epub 2015 Oct 7.
Epidermal growth factor (EGF) is an important factor for healing after tissue damage in diverse experimental models. It plays an important role in liver regeneration (LR). The objective of this experiment is to investigate the methylation variation of 10 CpG sites in the Egf promoter region and their relevance to Egf expression during rat liver regenera- tion. As a follow up of our previous study, rat liver tissue was collected after rat 2/3 partial hepatectomy (PH) during the re-organization phase (from days 14 to days 28). Liver DNA was extracted and modified by sodium bisulfate. The methylation status of 10 CpG sites in Egf promoter region was determined using bisulfite sequencing polymerase chain reaction (PCR), as BSP method. The results showed that 3 (sites 3, 4 and 9) out of 10 CpG sites have strikingly methylation changes during the re-organization phase compared to the regeneration phase (from 2 hours to 168 hours, P=0.002, 0.048 and 0.018, respectively). Our results showed that methylation modification of CpGs in the Egf promoter region could be restored to the status before PH operation and changes of methylation didn't affect Egf mRNA expression during the re-organization phase.
表皮生长因子(EGF)是多种实验模型中组织损伤后愈合的重要因子。它在肝脏再生(LR)中起重要作用。本实验的目的是研究大鼠肝脏再生过程中Egf启动子区域10个CpG位点的甲基化变化及其与Egf表达的相关性。作为我们先前研究的后续,在大鼠2/3部分肝切除(PH)后的重组阶段(第14天至第28天)收集大鼠肝脏组织。提取肝脏DNA并用亚硫酸氢钠进行修饰。使用亚硫酸氢盐测序聚合酶链反应(PCR),即BSP方法,测定Egf启动子区域10个CpG位点的甲基化状态。结果显示,与再生阶段(2小时至168小时)相比,10个CpG位点中有3个(位点3、4和9)在重组阶段有显著的甲基化变化(P分别为0.002、0.048和0.018)。我们的结果表明,Egf启动子区域CpG的甲基化修饰可恢复到PH手术前的状态,并且在重组阶段甲基化的变化不影响Egf mRNA表达。