Research Center for Traditional Chinese Medicine Complexity System, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Department of Medical Oncology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Virol Sin. 2019 Oct;34(5):489-500. doi: 10.1007/s12250-019-00117-0. Epub 2019 Jun 3.
The study was conducted to explore the mechanisms of sex differences in the response to chronic hepatitis B (CHB) in terms of DNA methylation, SNP genotype, and gene expression. Genomic DNA was isolated from peripheral blood mononuclear cells (PBMCs) of CHB patients and healthy controls and evaluated using the Human Methylation 450 K Assay. The DNA methylation level at hg37 chromosome (CHR) X: 7810800 was further validated using pyrosequencing. SNP genotypes, VCX mRNA expression of PBMCs, and plasma VCX protein concentration were further examined using SNaPshot, RT-qPCR, and Western blot, respectively. Results showed that a total of 5529 CpG loci were differentially methylated between male and female CHB patients. DNA methylation level and CC + CT frequency at CHR X: 7810800, VCX mRNA expression of PBMCs, and plasma VCX protein concentration were higher in female than in male CHB patients. The CHR X: 7810800 locus was hypermethylated in CHB patients with CC + CT genotypes in comparison with those with the TT genotype. In cases of CC + CT genotypes, VCX mRNA expression was negatively correlated with the DNA methylation level. CHB patients with higher levels of HBV DNA, AST, and GGT or higher GPRI scores exhibited lower VCX expression. In conclusion, SNPs and DNA methylation at the CHR X: 7810800 locus cooperatively regulate VCX expression in CHB. The upregulated VCX expression in female CHB patients might represent a mechanism of protection from more severe liver dysfunction and extensive fibrosis, as observed in male CHB patients.
该研究旨在探讨慢性乙型肝炎（CHB）患者中性别差异对 DNA 甲基化、SNP 基因型和基因表达的影响机制。从 CHB 患者和健康对照者的外周血单核细胞（PBMC）中分离基因组 DNA，并使用 Human Methylation 450 K Assay 进行评估。进一步使用焦磷酸测序验证 hg37 染色体（CHR）X：7810800 处的 DNA 甲基化水平。进一步使用 SNaPshot、RT-qPCR 和 Western blot 分别检测 SNP 基因型、PBMC 中 VCX mRNA 表达和血浆 VCX 蛋白浓度。结果表明，男性和女性 CHB 患者之间共有 5529 个 CpG 位点存在差异甲基化。女性 CHB 患者的 DNA 甲基化水平和 CHR X：7810800 处 CC+CT 频率、PBMC 中 VCX mRNA 表达和血浆 VCX 蛋白浓度均高于男性 CHB 患者。与 TT 基因型相比，CC+CT 基因型的 CHB 患者 CHR X：7810800 位点的甲基化程度更高。在 CC+CT 基因型的情况下，VCX mRNA 表达与 DNA 甲基化水平呈负相关。HBV DNA、AST 和 GGT 水平较高或 GPRI 评分较高的 CHB 患者表现出较低的 VCX 表达。总之，CHR X：7810800 位点的 SNP 和 DNA 甲基化共同调节 CHB 中的 VCX 表达。女性 CHB 患者中 VCX 表达的上调可能代表了一种保护机制，可避免男性 CHB 患者出现更严重的肝功能障碍和广泛纤维化。
