硒蛋白基因DIO2启动子区CpG甲基化与CpG-SNPs的交互作用对大骨节病的影响

doi:10.24920/003953

摘要/Abstract

摘要：

目的本研究旨在明确大骨节病（KBD）患者硒蛋白DIO2基因启动子区4个CpGs的甲基化水平和2个CpG-SNPs的基因型,并探讨两者的相关性。
方法采用基质辅助激光解吸电离飞行时间质谱技术（MALDI-TOF-MS）检测KBD患者（n=16）和健康对照组（n=16）血液中DIO2基因启动子区的4个CpGs和2个CpG-SNPs,分析比较两组及不同CpG-SNP基因型KBD患者CpGs甲基化水平。
结果 KBD患者全血中DIO2 mRNA水平明显低于健康对照组（P<0.05）,KBD患者DIO2-1_CpG_3甲基化水平明显高于健康对照组（P<0.05）,4个CpGs的甲基化水平在KBD患者和健康对照组无明显差异（P>0.05）,GA/AA基因型（DIO2 rs955849187）KBD患者DIO2启动子区DIO2-1_CpG_3甲基化水平显著高于GG基因型KBD患者（P<0.05）。
结论 KBD患者DIO2启动子区甲基化水平升高,CpG-SNPs DIO2 rs955849187变异基因型的KBD患者DIO2启动子区甲基化水平亦呈升高趋势。

关键词: 大骨节病, DNA甲基化, CpG位点, CpG-SNP

Abstract:

Objective This study was designed to determine the methylation profile of four CpGs and the genotypes of two CpG-SNPs located in promoter region of DIO2 in patients with Kashin-Beck disease (KBD). We also analyzed the interaction between the CpGs methylations and CpG-SNPs.
Methods Whole blood specimens were collected from 16 KBD patients and 16 healthy subjects. Four CpGs and two CpG-SNPs in the promoter regions of DIO2 were detected using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The CpGs methylation levels were compared between samples from KBD patients and healthy subjects. The methylation levels were also analyzed in KBD patients with different CpG-SNP genotypes.
Results The mRNA expression of DIO2 in whole blood of KBD patients was significnatly lower than in healthy controls (P <0.05). The methylation levels of DIO2-1_CpG_3 in KBD patients were significantly higher than those in healthy controls (P <0.05). The methylation levels of four CpGs were not significantly different between KBD patients and healthy controls. The methylation level of DIO2-1_CpG_3 in the promoter region of DIO2 in KBD patients with GA/AA genotype was significantly higher than that of KBD patients with GG genotype (P <0.05).
Conclusion The methylation level of DIO2 increases in KBD patients. Similar trends exist in KBD carriers of variant genotypes of CpG-SNPs DIO2 rs955849187.

Key words: Kashin-Beck disease, DNA methylation, CpG, CpG-SNP

基金资助: 国家自然科学基金(82073494);陕西省重点研发计划(2020SF-076);中国富硒产业研究院富硒产业全产业链专项研发计划项目(2020FXZX0501);陕西中医药大学重点创新团队(132041933)

Rongqiang Zhang, Dandan Zhang, Di Zhang, Xiaoli Yang, Qiang Li, Chen Wang, Xuena Yang, Yongmin Xiong. Crosstalk between CpG Methylation and Polymorphisms (CpG-SNPs) in the Promotor Region of DIO2 in Kashin-Beck Disease[J].Chinese Medical Sciences Journal, 2022, 37(1): 52-59.

图/表 8

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CpG-SNPs	Primer Sequence		Tm(°C)
rs914454577	Fw:	ACGTTGGATGGTATAAAAGGCGTCAGGGAG	48.6
rs914454577	Rv:	ACGTTGGATGTTGCTATCTGTCTGTGGTGC	48.6
rs955849187	Fw:	ACGTTGGATGGAGCGACTGACTCCTCTTTT	50.8
rs955849187	Rv:	ACGTTGGATGCAGAGAGAGGCACTTTGCAC	50.8

Characteristics	KBD (n=16)	Controls (n=16)	t/x²	P value
Age, $\bar{x} \pm SD$	54.26±5.61	52.35±8.79	0.733	0.469
Gender, F/M	6/10	8/8	0.508	0.476
Degree, n(%)
I	1 (6.25)	-
II	7 (43.75)	-
III	8 (50.00)	-

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