Clinical Laboratory of Renmin Hospital, The Third Affiliated Hospital of Hubei University of Medicine, 39 Chaoyang Road, Shiyan, Hubei Province 442000, China.
J Diabetes Res. 2013;2013:721493. doi: 10.1155/2013/721493. Epub 2013 Mar 16.
Objectives. To study the correlation between the methylation of protein kinase C epsilon zeta (PRKCZ) gene promoters and type 2 diabetes mellitus (T2DM). Methods. The case-control method was implemented in 272 unrelated to one another cases in Shiyan People's Hospital. Of those, 152 were diagnosed as T2DM cases, and the other 120 cases were healthy individuals visiting the hospital for a physical examination. The subjects were first divided into two groups: the T2DM group and the normal control (NC) group. Next, methylated DNA immunoprecipitation chip (MeDIP-chip) was used for detection. Bisulfite sequencing PCR (BSP) and gene sequencing were then performed to detect and analyze the correlation between PRKCZ gene promoter methylation and T2DM. Finally, Western blotting was applied to determine the serum level of PRKCZ. The data were then analyzed with the statistics analyzing software SPSS 17.0. Results. In contrast with cases in NC, T2DM patients showed a high level of methylation, with 7 of 9 CpG sites were shown to be methylated, whereas, in the control group, only one CpG site was found to be methylated. The methylated CpG sites for the two groups showed marked differences (P < 0.01). Additionally, the level of PRKCZ was decreased in T2DM subjects, and the difference between the two groups was statistically significant (P < 0.05). Discussion. This study suggests that the PRKCZ gene is the hypermethylated gene of T2DM and the hypermethylation PRKCZ gene may be involved in the pathogenesis of T2DM.
目的。研究蛋白激酶 C epsilon zeta(PRKCZ)基因启动子甲基化与 2 型糖尿病(T2DM)的相关性。方法。采用病例对照方法,对十堰人民医院的 272 例非相关病例进行研究。其中,152 例被诊断为 T2DM 病例,120 例为健康体检者。将这些受试者首先分为两组:T2DM 组和正常对照组(NC)。然后使用甲基化 DNA 免疫沉淀芯片(MeDIP-chip)进行检测。接着进行亚硫酸氢盐测序 PCR(BSP)和基因测序,以检测和分析 PRKCZ 基因启动子甲基化与 T2DM 的相关性。最后,应用 Western 印迹法测定 PRKCZ 血清水平。使用统计分析软件 SPSS 17.0 对数据进行分析。结果。与 NC 组相比,T2DM 患者的甲基化水平较高,9 个 CpG 位点中有 7 个被发现甲基化,而在对照组中,只有一个 CpG 位点被发现甲基化。两组的甲基化 CpG 位点存在明显差异(P < 0.01)。此外,T2DM 患者的 PRKCZ 水平降低,两组间差异具有统计学意义(P < 0.05)。讨论。本研究表明,PRKCZ 基因是 T2DM 的高甲基化基因,高甲基化 PRKCZ 基因可能参与 T2DM 的发病机制。