Research Paper Volume 13, Issue 4 pp 4831—4849
An aging and p53 related marker: HOXA5 promoter methylation negatively correlates with mRNA and protein expression in old age
- 1 Department of General-, Visceral-, Vascular- and Pediatric Surgery, University of Saarland Medical Center, Homburg 66421, Saar, Germany
- 2 Department of Genetics and Epigenetics, Saarland University, Saarbrücken 66123, Germany
Received: September 10, 2020 Accepted: January 4, 2021 Published: February 5, 2021https://doi.org/10.18632/aging.202621
How to Cite
Copyright: © 2021 Feiner et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
The process of aging has been associated with differential patterns of DNA methylation which relate to changes in gene expression. Hence, we aimed to identify genes with significant age-related methylation differences and to study their mRNA and protein expression profile. Genome-wide DNA methylation analysis was performed with the Illumina Infinium Methylation EPIC BeadChip Microarray on bisulfite-converted DNA prepared from monocytes derived from young (average age: 23.8 yrs) and old (average age: 81.5 yrs) volunteers that are separated by at least 50 years of age difference, n=4, respectively. Differentially methylated CpG sites were assigned to the associated genes and validated by deep sequencing analysis (n=20). Demonstrating an age-associated significant increase of methylation in the promoter region (p=1x10-8), Homeobox A5 (HOXA5), also known to activate p53, emerged as an interesting candidate for further expression analyses by Realtime PCR, ELISA and Western Blot Analysis (n=30, respectively). Consistent with its hypermethylation we observed significant HOXA5 mRNA downregulation (p=0.0053) correlating with significant p53 downregulation (p=0.0431) in the old cohort. Moreover, we observed a significant change in HOXA5 protein expression (p=3x10-5) negatively correlating with age and promoter methylation thus qualifying HOXA5 for an eligible p53-related aging marker.
HOXA5: Homeobox protein A5; CpG: Cystein phosphatidyl Guanin; p53: Phosphoprotein p53; ELISA: Enzyme-linked immunosorbent assay; PBMC: Peripheral blood mononuclear cells.