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Evaluation of DNMT1 gene expression profile and methylation of its promoter region in patients with ankylosing spondylitis

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Abstract

Ankylosing spondylitis (AS) is an autoimmune disease with a chronic inflammatory arthritis. The critical role of methylation in the biology of immunocytes has increasingly been surveyed to discover disease etiology. DNA methyltransferase 1 (DNMT1) is an enzyme, which establishes and regulates patterns of methylated cytosine residues. The aim of the current investigation was to unveil if methylation circumstances of CpG sites in DNMT1 promoter could affect the mRNA expression level of this gene in peripheral blood mononuclear cells (PBMCs) from AS patients. PBMCs were isolated from whole blood of 40 AS patients and 40 healthy individuals. Total RNA and DNA contents of leukocytes were extracted. Afterward, quantitative analysis was carried out by real-time PCR using the SYBR Green PCR Master Mix. Finally, to determine the methylation level, PCR products of bisulfite-treated DNA from patients and controls were sequenced. Compared with healthy controls, expression level of DNMT1 in AS patients was significantly downregulated. Methylation of DNMT1 promoter was significantly higher in AS patients in comparison to controls. While a negative correlation between methylation and expression level of DNMT1 was observed in AS patients, both methylation and expression level of DNMT1 did not correlate with clinical manifestations. Considering the observation that decreased expression level of DNMT1 was associated with hypermethylation of DNMT1 promoter in PBMCs from AS patients, this survey suggests that dysregulation of DNMT1 expression through altered methylation level of other target genes would probably contribute to AS development.

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Acknowledgments

We are deeply grateful to our participants for their contribution in the accomplishment of this investigation. The work reported herein was supported by a grant from Deputy of Research, Tehran University of Medical Sciences (Grant No. 93-03-03-27132).

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Authors and Affiliations

  1. Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Saeed Aslani, Jafar Karami & Mohammad Hossein Nicknam

  2. Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran

    Mahdi Mahmoudi & Ahmad Reza Jamshidi

  3. Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Masoud Garshasbi

  4. Molecular Immunology Research Center, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Mohammad Hossein Nicknam

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  1. Saeed Aslani
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  2. Mahdi Mahmoudi
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  3. Masoud Garshasbi
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  4. Ahmad Reza Jamshidi
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  5. Jafar Karami
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  6. Mohammad Hossein Nicknam
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Corresponding authors

Correspondence to Mahdi Mahmoudi or Mohammad Hossein Nicknam.

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The Human Research Ethics Committee from the Tehran University of Medical Sciences approved this study. Written informed consent was taken by all the participants.

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Aslani, S., Mahmoudi, M., Garshasbi, M. et al. Evaluation of DNMT1 gene expression profile and methylation of its promoter region in patients with ankylosing spondylitis. Clin Rheumatol 35, 2723–2731 (2016). https://doi.org/10.1007/s10067-016-3403-x

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  • DOI: https://doi.org/10.1007/s10067-016-3403-x

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