摘要
目的研究盘状红斑狼疮(DLE)患者T淋巴细胞CD70 mRNA的表达及CD70基因启动子DNA甲基化的状态,探讨CD70在DLE发病机制中作用。方法将2014年1月至2015年6月的15例活动期、15例非活动期DLE患者和15例正常人对照组外周血中T淋巴细胞的分离以及DNA、RNA提取,然后以定量RT-PCR测定CD4^+细胞和CD8^+细胞中CD70 mRNA转录水平,亚硫酸氢钠基因测序法检测CD4^+细胞和CD8^+细胞CD70基因启动子区域甲基化水平。结果活动期、非活动期和健康对照组DLE患者CD4^+T淋巴细胞中的CD70 mRNA转录水平分别为(0.82±0.21)、(0.61±0.15)和(0.43±0.11),呈逐步递减趋势,且两两比较差异均有显著统计学意义(P<0.01),活动期、非活动期DLE患者和健康对照CD4^+T淋巴细胞CD70基因启动子序列-600^-300 bp区域平均甲基化水平分别为(0.29±0.05)、(0.41±0.03)和(0.59±0.02),呈逐步递增趋势,且两两比较差异均有统计学意义(P<0.01).结论 DLE患者T淋巴细胞中CD4^+细胞CD70基因启动子区域的低甲基化状态可能引起CD70过度表达,这为DLE疾病患者的检测提供又一指标。
Objective To detect the expressions of CD70 mRNA, determine the methylation status of CD70 gene promoter in T cells from patients with discoid lupus erythematosus(DLE), and explore the role of CD70 in mechanism of DLE. Methods Blood samples were obtained from 15 patients with active DLE, 15 patients with unreactive DLE and 15 normal human controls from January 2014 to June 2015. Quantitative RT-PCR was carried out to quantify the mRNA expression of CD70, and bisulfite sequencing was used to evaluate the methylation status of CD70 gene promoter in CD4^+and CD8^+T cells. Results The transcriptional level of CD70 mRNA in CD4^+T lymphocytes in active DLE patients, inactive patients and healthy controls were respectively(0.82±0.21),(0.61±0.15) and(0.43±0.11), with a decreasing trend. There are significant differences in each paired comparison(P<0.01). The average methylation index of the region between-600 bp and-300 bp of CD70 gene promoter in CD4^+T cells of the patients with active DLE(0.29 ± 0.05) and inactive DLE(0.41±0.03), which was significantly lower than that in the healthy controls(0.59±0.02)(P<0.01). Conclusion The CD70 gene promoter in CD4^+T cells is significantly hypomethylated in patients with DLE, which may directly lead to the overexpression of CD70, and could be used as an important index of disease detection in patients with DLE.
出处
《海南医学》
CAS
2016年第13期2070-2072,2073,共4页
Hainan Medical Journal
基金
海南省卫生厅卫生计生行业科研项目(编号:琼卫2013自筹-33)
