KIR3 DL1启动子区域 CpG 岛甲基化对抗原表达的影响
Effects of methylated CpG islands in promoter region on the expression of KIR3DL1 protein
摘要目的:探索KIR3DL1启动子区域CpG岛甲基化对抗原表达的影响。方法选择抗原高表达KIR3DL1倡01502和低表达KIR3DL1倡005样本,分别测定启动子区域碱基序列和启动子区CpG岛甲基化程度。对携带有KIR3DL1倡01502、KIR3DL1倡005的NK细胞分别采用5-aza进行去甲基化处理,利用流式细胞仪检测KIR3DL1抗原。结果 KIR3DL1倡01502和KIR3DL1倡005启动子区域-65和-269位存在碱基差异,这导致它们的启动子区域有两个不同的CpG岛。 KIR3DL1倡01502启动子区CpG岛高度甲基化,去甲基化后相应的细胞表面抗原表达显著增加。而携带KIR3DL1倡005的细胞去甲基化处理后,抗原表达没有明显变化。结论 KIR3DL1倡01502启动子区域CpG岛甲基化影响抗原的表达,但是不同抗原表达模式的KIR3DL1等位基因可能存在不同的调控机制。
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abstractsObjective To investigate the effects of methylated CpG islands in the promoter region on the expression of killer cell immunoglobulin-like receptor 3DL1 (KIR3DL1).Methods Three voluntary unpaid blood donors carrying high expression allele KIR 3DL1*01502 and three donors carrying low expres-sion allele KIR3DL1*005 were recruited in this study .The nucleotide sequences and the methylated CpG islands in the promoter regions of KIR 3DL1*01502 allele and KIR3DL1*005 allele were analyzed .The NK cells expressing KIR3DL1*01502 and KIR3DL1*005 were respectively treated with 5-aza for the dem-ethylation of CpG islands within the promoters .The expression of KIR3DL1 protein on the surface of NK cells was measured with flow cytometer .Results Two differences at nucleotide sites -65 and -269 were detected within the promoter regions of KIR3DL1*01502 and KIR3DL1*005, resulting in two distinct CpG islands.The CpG islands within the promoter of KIR 3DL1*01502 allele were highly methylated .The ex-pression of KIR3DL1 protein on NK cells which carried KIR 3DL1*01502 allele was significantly increased after the demethylation of CpG islands .However , the treatment of demethylation had no significant effects on the expression KIR3DL1 protein on NK cells harboring KIR3DL1*005 allele.Conclusion The methylated CpG islands within the promoter of KIR 3DL1*01502 allele affected the antigen expression on the surface of NK cells.Different KIR3DL1 alleles might show different mechanisms in regulating antigen expression .
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