中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (2): 80-83.

• 论著 • 上一篇    下一篇

系统性红斑狼疮患者T淋巴细胞CD70基因表达及其启动子区域甲基化状态的研究

丁艳1,肖嵘2,张燕蒙秉新4,苏建英5,韩科5   

  • 收稿日期:2012-04-09 修回日期:2012-10-10 出版日期:2013-02-15 发布日期:2013-02-01
  • 通讯作者: 丁艳 E-mail:annymilk@126.com
  • 基金资助:
    海南省自然科学基金

Expressions of CD70 mRNA and protein and methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus

  • Received:2012-04-09 Revised:2012-10-10 Online:2013-02-15 Published:2013-02-01
  • Supported by:
    Natural Science Foundation of Hainan Province of China

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摘要： 目的 探讨系统性红斑狼疮（SLE）患者T淋巴细胞CD70mRNA和蛋白的表达及CD70基因启动子DNA甲基化的状态。方法 分离15例活动期SLE患者、15例非活动期SLE患者和15例健康对照外周血CD4+与CD8+细胞，用实时定量逆转录PCR（RT-PCR）方法检测CD4+和CD8+细胞CD70 mRNA转录水平，流式细胞仪检测CD4+CD70+ 细胞和CD8+CD70+细胞百分率，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。组间比较采用单因素方差分析，组间两两比较采用SNK-q检验。 结果 ①活动期、非活动期SLE患者CD4+细胞CD70 mRNA转录水平分别为0.82 ± 0.12和0.73 ± 0.11，明显高于健康对照组（0.45 ± 0.09），F = 53.017，P < 0.01，活动期SLE患者CD4+细胞的CD70 mRNA转录水平显著高于非活动期SLE患者（P < 0.05）。活动期、非活动期SLE患者外周血CD4+CD70+细胞百分率分别为80.30% ± 11.04% 和66.80% ± 3.98%，明显高于健康对照组（12.48% ± 3.45%），F = 311.517，P < 0．01，活动期SLE患者CD4+CD70+细胞百分率显著高于非活动期SLE患者（P值 < 0.05）。SLE患者外周血CD70+CD4+细胞百分率与SLE疾病活动度呈显著正相关（r = 0.792，P = 0.000）。活动期、非活动期SLE患者组的CD4+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0.32 ± 0.05和0.36 ± 0.05，明显低于健康对照组（0.62 ± 0.05），F = 152.64，P < 0.01，活动期平均甲基化水平明显低于非活动期SLE患者组（P < 0.05）。结论 CD4+细胞CD70基因启动子区域处于低甲基化状态，这种低甲基化状态可能是CD70过度表达的直接原因。

关键词: 红斑狼疮，系统性, T淋巴细胞, 抗原 CD70, DNA甲基化

Abstract: DING Yan*, XIAO Rong, ZHANG Yan, MENG Bing-xin, SU Jian-ying, HAN Ke. *Department of Dermatology and Venereology, Hainan Provincial People's Hospital, Haikou 570102, China 【Abstract】 Objective To detect the expressions of CD70 mRNA and protein and to determine the methylation status of CD70 gene promoter in T cells from patients with systemic lupus erythematosus（SLE）．Methods Peripheral blood CD4+ and CD8+ T cells were isolated from 15 patients with active SLE, 15 patients with inactive SLE and 15 healthy control subjects． Real-time quantitative reverse transcription-PCR was carried out to quantify the mRNA expression of CD70, flow cytometry to determine the frequency of CD4+CD70+ and CD8+CD70+ T cells, and bisulfite sequencing to evaluate the methylation status of CD70 gene promoter in CD4+ and CD8+ T cells． Differences in these parameters among these groups were analyzed by one-factor analysis of variance and SNK-q test. Results Compared with the healthy controls, the patients with active SLE and inactive SLE showed a significant increase in CD70 mRNA expression in CD4+ T cells （0.82 ± 0.12 and 0.73 ± 0.11 vs. 0.45 ± 0.09, F = 53.017, P < 0.01） and in the frequency of CD70+CD4+ T cells （80.30% ± 11.04% and 66.80% ± 3.98% vs. 12.48% ± 3.45%, F = 311.517, P < 0.01）. Also, the expression of CD70 mRNA in CD4+ T cells and the frequency of CD70+CD4+ T cells were significantly higher in patients with active SLE than in patients with inactive SLE （both P < 0.05）. There was a positive correlation between the frequency of peripheral CD70+CD4+ T cells and disease activity in SLE in these patients（r = 0.792, P < 0.01）. The average methylation index of the region between －600 bp and －300 bp of CD70 gene promoter in CD4+ T cells was 0.32 ± 0.05 and 0.36 ± 0.05 respectively in the patients with active and inactive SLE, significantly lower than that in the healthy controls （0.62 ± 0.05, F = 152.64, P < 0.01）, and the patients with active SLE showed a significantly lower methylation index than those with inactive SLE （P < 0.05）. Conclusions The CD70 gene promoter in CD4+ T cells is significantly hypomethylated in patients with SLE, which may directly lead to the overexpression of CD70. 【Key words】 Lupus erythematosus, systemic; T-lymphocytes; Antigens, CD70; DNA methylation

Key words: T-lymphocytes, Antigens，CD70, DNA methylation