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Allele-specific methylation contributed by CpG-SNP is associated with regulation of ALOX5AP gene expression in ischemic stroke

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Abstract

Previous studies have shown that CpG-SNPs might have influence on gene function via allele-specific DNA methylation (ASM). However, association study between DNA methylation and the promoter CpG-SNPs in ALOX5AP gene with IS has not been reported. The present study aims to explore the relationship among CpG-SNPs, methylation levels, and messenger RNA (mRNA) expression levels of ALOX5AP gene. Firstly, we made a two-stage association study to identify a potential associated CpG-SNP (rs4073259) by SNaPshot genotyping approach (P = 0.015, OR = 0.672, 95% CI 0.487–0.927; P = 0.035, OR = 0.809, 95% CI 0.664–0.985, respectively). In addition, the methylation levels of 17 CpG sites located in the promoter of ALOX5AP were tested by MethylTarget sequencing. The methylation level of GG genotype carriers is significantly higher than those with the AG and AA genotypes (P < 0.05). And the GG genotype carriers with higher DNA methylation levels have a decreased mRNA expression levels of ALOX5AP (P < 0.05). Finally, we found that the G allele with higher methylation level has got a lower transcription activity than the A allele by luciferase assay (P = 0.000).The study provided evidence that IS-associated CpG-SNP rs4073259 may affect the expression level of ALOX5AP through allele-specific methylation and consequently the phenotype of the disease.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (No. 81571154) and by the National Natural Science Foundation of China (No. 81671163). We would like to thank the technical assistance of staff members of the Neurology Department of the First Affiliated Hospital of Zhengzhou University and the First People Hospital of Zhengzhou. We also thank all the subjects providing blood samples.

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  1. Yunshu Shi and Liyan Xu contributed equally.

Authors and Affiliations

  1. Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China

    Yunshu Shi

  2. Department of Medical Genetics and Cell Biology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China

    Liyan Xu, Qingchuan Feng, Jing Jia, Yan Xu, Xuran Zhang, Huiling Zhao, Jun Wang, Ying He & Hong Zheng

  3. Department of Neurology, The First People’s Hospital of Zhengzhou, Zhengzhou, 450004, China

    Aifan Li

  4. School of Life Sciences of Zhengzhou University, Zhengzhou, 450001, China

    Dongzhi Yang

  5. Department of Eugenic Genetics, The First People’s Hospital of Zhengzhou, Zhengzhou, 450004, China

    Yuchao Zhang

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  1. Yunshu Shi
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Contributions

Y.S.S., Y.C.Z., and H.L.Z. performed the experiments. J.J., D.Z.Y., and Y.X. conducted the statistical analysis. A.F.L., Q.C.F., X.R.Z., and J.W. were involved in the participant recruitment and local study implementation. Y.H. and L.Y.X. wrote the manuscript. Y.H. and H.Z. conceived of the study, and participated in its design and coordination. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Ying He or Hong Zheng.

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Shi, Y., Xu, L., Feng, Q. et al. Allele-specific methylation contributed by CpG-SNP is associated with regulation of ALOX5AP gene expression in ischemic stroke. Neurol Sci 39, 1717–1724 (2018). https://doi.org/10.1007/s10072-018-3489-9

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