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Global DNA methylation profile at LINE-1 repeats and promoter methylation of genes involved in DNA damage response and repair pathways in human peripheral blood mononuclear cells in response to γ-radiation

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Abstract

DNA methylation is an epigenetic mechanism, which plays an important role in gene regulation. The present study evaluated DNA methylation profile of LINE1 repeats and promoter methylation of DNA damage response (DDR) and DNA repair (DR) genes (PARP1, ATM, BRCA1, MLH1, XPC, RAD23B, APC, TNFα, DNMT3A, MRE11A, MGMT, CDKN2A, MTHFR) in human peripheral blood mononuclear cells (PBMCs) of healthy donors in response to γ-radiation. Methylation level was correlated with gene expression profile of selected DDR and DR genes (APC, MLH1, PARP1, MRE11A, TNFα, MGMT) to understand their role in gene regulation. Blood samples were collected from 15 random healthy donors, PBMCs were isolated, exposed to 0.1 Gy (low) and 2.0 Gy (high) doses of γ-radiation and proliferated for 48 h and 72 h. Genomic DNA and total RNA were isolated from irradiated PBMCs along with un-irradiated control. Methylation profile was determined from bisulphite converted DNA and amplified by methylation sensitive high resolution melting (MS-HRM) method. Total RNA was converted to cDNA and relative expression was analysed using real time quantitative-PCR. Our results revealed that at 0.1 Gy, MRE11A and TNFα showed significant (P < 0.05) increase in methylation at 72 h. At 2.0 Gy, significant increase (P < 0.05) in methylation profile was observed at LINE1, MRE11A, PARP1, BRCA1, DNMT3A and RAD23B at 48 h and 72 h. PARP1 showed significant positive correlation of methylation status with gene expression. In conclusion, low and high doses of γ-radiation have significant influence on DNA methylation status of LINE1, DDR and DR genes suggesting their potential role as epigenetic signatures in human PBMCs, which can be further explored in human populations.

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Abbreviations

IR:

Ionizing radiation

HDIR:

High doses of ionizing radiation

LDIR:

Low doses of ionizing radiation

LNT:

Linear No Threshold

LET:

Linear Energy Transfer

DDR:

DNA damage response

DR:

DNA repair

MS HRM:

Methyl Sensitive High Resolution Melting

PBMCs:

Peripheral Blood Mononuclear Cells

BRIT:

Board of Radiation and Isotope Technology

PHA:

Phytohaemagglutinin

EDTA:

Ethylene diamine tetra-acetic acid

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Acknowledgements

We are very much thankful to the volunteers, who have participated in the study. We profusely thank Mr. P.K.M. Koya for helping us in the statistical analysis. We also thank to the pathology staff members from BARC dispensary, Trombay for helping us in collecting the blood samples for this study.

Funding

The research work is funded by Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.

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Authors and Affiliations

  1. Low Level Radiation Research Section, Radiation Biology and Health Sciences Division, Bio-Sciences Group, Bhabha Atomic Research Centre, Trombay, Mumbai, 400 085, India

    Rashmi Priya & Birajalaxmi Das

  2. Homi Bhabha National Institute, Anushaktinagar, Trombay, Mumbai, 400 094, India

    Birajalaxmi Das

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  1. Rashmi Priya
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BD Conceived the concept, designed the experiment, implemented the work, analysed and interpreted the data, written and reviewed the manuscript. RP conducted the experiment, analyzed the data and involved in writing the initial draft of the manuscript.

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Correspondence to Birajalaxmi Das.

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All the volunteers have given written informed consent to participate in the study. All experiments were conducted in accordance with the ethical guidelines of the Medical Ethic Committee, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.

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Priya, R., Das, B. Global DNA methylation profile at LINE-1 repeats and promoter methylation of genes involved in DNA damage response and repair pathways in human peripheral blood mononuclear cells in response to γ-radiation. Mol Cell Biochem 477, 267–281 (2022). https://doi.org/10.1007/s11010-021-04265-4

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