TNFRSF1A基因CpG岛甲基化与基因表达及T淋巴细胞亚群指标的相关性分析

doi:10.11843/j.issn.0366-6964.2017.10.004

摘要/Abstract

摘要：

旨在探讨TNFRSF1A基因CpG岛的甲基化水平，阐明该基因甲基化与基因表达及T淋巴细胞亚群性状的关系。本研究以抗病力强的华北型黑猪大蒲莲猪和抗病力相对较弱的引进猪种长白猪的仔猪为试验群体，通过亚硫酸氢盐处理后测序法（BSP）探求TNFRSF1A基因上游CpG岛甲基化情况，并将甲基化位点的甲基化频率与基因表达量及T淋巴细胞亚群性状进行相关性分析。结果表明：1） TNFRSF1A基因起始位点附近预测到2个CpG岛（Island），跨这2个CpG岛进行BSP测序后共发现25个CpG位点，其中Island 1和Island 2中各有9个CpG位点，其余位于2个CpG岛之间。Island 1的甲基化频率在大蒲莲和长白猪两群体间差异不显著（P>0.05），Island 2的甲基化频率大蒲莲猪群体显著高于长白猪群体（P<0.05）。2）Island 2中甲基化频率与ΔCt值成正相关，即甲基化频率与基因表达量成负相关，其中+126、+128、+159、+162和+164是重要的甲基化位点。3）两个猪群体中位于Island 2的+159和+164位点与CD4+CD8+CD3+指标显著正相关（P<0.05）。Island 2中+159和+164位点甲基化频率与基因表达量成负相关，而且这2个位点又与T淋巴细胞亚群CD4+CD8+CD3+显著正相关，笔者推测，大蒲莲仔猪TNFRSF1A基因+159和+164位点的甲基化频率增加，可能导致TNFRSF1A基因表达量下降，影响T淋巴细胞亚群性状，从而影响机体的免疫水平。

Abstract:

The aim of this study was to explore CpG islands methylation status of TNFRSF1A gene, and clarify the relationship of methylation level with its expression and T lymphocyte subpopulation traits. In the present study, Dapulian piglets, one indigenous breed with higher disease resistance in North China, and Landrace piglets, one commercial breed with lower disease resistance, were used. Methylation level of CpG islands of TNFRSF1A gene upstream was determined by bisulfite sequencing PCR (BSP), and correlation analyses of methylation frequency of CpG sites with its expression and T lymphocyte subpopulation traits were carried out. The results indicated that: 1) There were 2 CpG islands in the upstream of TNFRSF1A gene, which contained 25 CpG sites by BSP overlapping 2 islands. Nine CpG sites were in Island 1, nine CpG sites were in Island 2, and others located between 2 islands. Population methylation frequency between Dapulian and Landrace piglets had no significant difference in Island 1 at 0.05 level, whereas methylation frequency of Dapulian piglets was significantly higher than that of Landrace piglets in Island 2 at 0.05 level. 2) The methylation frequencies in Island 2 were negatively correlated with the expression of TNFRSF1A, especially 5 CpG sites (+126, +128, +159, +162 and +164). 3) We also found methylation frequencies of CpG sites +159 and +164 were positively correlated with CD4+CD8+CD3+ at 0.05 level in the 2 populations. The methylation frequencies of CpG sites +159 and +164 in Island 2 were negatively correlated with the expression of TNFRSF1A, and positively correlated with T lymphocyte subpopulation trait CD4+CD8+CD3+. Thus, we speculated that increased methylation frequencies of CpG sites +159 and +164 might lead to the decreased expression of TNFRSF1A, which may influence on T lymphocyte subpopulation traits and the immunity of Dapulian piglets.

中图分类号:

S828.2

赵雪艳, 王彦平, 王怀中, 郭建凤, 齐波, 王继英. TNFRSF1A基因CpG岛甲基化与基因表达及T淋巴细胞亚群指标的相关性分析[J]. 畜牧兽医学报, 2017, 48(10): 1815-1824.

ZHAO Xue-yan, WANG Yan-ping, WANG Huai-zhong, GUO Jian-feng, QI Bo, WANG Ji-ying. Correlation Analyses of CpG Island Methylation of TNFRSF1A with Gene Expression and T Lymphocyte Subpopulation Traits[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(10): 1815-1824.

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