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ITP中IFN-γ和IL-4 mRNA表达与其甲基化水平的相关性研究

赵海丰1,杨仁池2   

  1. 1. 天津医科大学附属肿瘤医院
    2. 中国医学科学院血液病医院
  • 收稿日期:2012-03-19 修回日期:2012-06-25 出版日期:2012-10-15 发布日期:2012-10-15
  • 通讯作者: 赵海丰

The mRNA Expression of IFN-γ and IL-4 are not Correlated with their DNA Methylation Status in ITP Patient

  • Received:2012-03-19 Revised:2012-06-25 Published:2012-10-15 Online:2012-10-15
  • Contact: Hai-Feng ZHAO

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摘要： 目的：检测免疫性血小板减少症(ITP)患者的干扰素（IFN）-γ和白细胞介素（IL）-4的mRNA的表达水平及其各自基因启动子区DNA甲基化的水平。方法：取36例ITP患者（ITP组）和36例正常对照外周血（对照组），采用实时定量PCR方法检测IFN-γ和IL-4 的mRNA水平的表达；随机抽取2组中的各10例，采用DNA甲基化修饰后测序的方法检测IFN-γ和IL-4基因启动子区的甲基化水平，并计算IFN-γ和IL-4基因的mRNA水平的表达量与其基因启动子区DNA甲基化水平的相关性。结果: 与对照组相比，ITP组IFN-γ的mRNA水平增高(1.86±0.29 vs 0.83±0.14, P< 0.05)，而IL-4 的mRNA水平降低 (0.78±0.22 vs 1.45±0.40, P< 0.05) ，Th1/Th2（IFN-γ/IL-4）比值升高 (7.11±0.60 vs 3.12±1.88, P< 0.01)。IFN-γ和IL-4基因CpG岛的位点甲基化水平和整体甲基化水平差异均无统计学意义（P >0.05）。ITP组中IFN-γ和IL-4的mRNA表达水平与基因启动子区甲基化水平不存在相关性（均P > 0.05）。结论： ITP患者处于Th1极化状态，IFN-γ和IL-4的基因启动子区的甲基化水平对各自基因mRNA水平的表达不起调控作用。

关键词: 血小板减少, 自身免疫疾病, 干扰素Ⅱ型, DNA甲基化, 白细胞介素4, RNA, 信使, DNA甲基化

Abstract: Objective: DNA methylation had been researched in many autoimmune diseases. Immune thrombocytopenia (ITP) is an organ-specific autoimmune disorder characterized by accelerated platelet destruction and bleeding symptoms. DNA methylation negatively regulates the gene expression. The aim of the current study was to investigate the relationship between the transcription levels of the IFN-γ and IL-4 and the methylation status of those gene promoters in ITP patients. Methods: The mRNA expression levels and DNA methylation of the IFN-γ and IL-4 were detected by the real-time quantitative PCR and bisulfite genomic sequencing, respectively. The relationship between CpG methylation and the mRNA levels of those genes were determined. Results: When compared with the healthy controls, the IFN-γ mRNA levels of ITP patients were significantly higher (1.86±0.29 vs 0.83±0.14, P<0.05). The levels of IL-4 mRNA transcripts were notably lower in ITP patients (0.78±0.22 vs 1.45±0.40, P<0.05). Most importantly, the Th1/Th2 (IFN-γ/IL-4) was remarkably higher in ITP patients than that of the controls (7.11±0.60 vs 3.12±1.88, p<0.01). There were no closely relationship between the mRNA levels of IFN-γ and IL-4 genes and the methylation status of those gene promoters. Conclusion: The ITP patients were in the Th1 polarization. The mRNA expression of IFN-γ and IL-4 were not associated with the DNA methylation status of those genes in ITP patients, indicating that the IFN-γ and IL-4 mRNA were not controlled by the DNA methylation of those genes.

Key words: thrombocytopenia autoimmune diseases interferon type iiinterleukin-4 RNA, messenger DNA methylation gene expression, DNA甲基化