Screening Of Abnormal DNA Methylation Genes In Patients With Acne Vulgaris

Objective Acne vulgaris is the most common inflammatory inflammatory follicle sebaceous gland unit disease in adolescents,and severe cases lead to appearance-impairment,depression,and suicidal tendencies.DNA methylation is involved in the pathogenesis of chronic inflammatory diseases,but no reports on DNA methylation in acne patients have been reported at home and abroad.By screening for abnormal methylation of whole-genome DNA in patients with moderate-to-severe acne,the abnormal DNA methylation sites and methylation levels are initially identified.To lay the foundation for further construction of the methylation map of acne patients,and to provide information for elucidating the genetic mechanism of acne.Methods 1.Select the back lesions and peripheral blood of 4 patients with moderate to severe acne vulgaris who have a family history in the outpatient clinic of Shenzhen Medical University of Southern Medical University,and the skin tissue of back in 2 healthy subjects was taken as control.The skin samples of patients and peripheral blood were matched with those of healthy controls.2.Whole body genomic DNA of skin tissue and peripheral blood samples were extracted using a DNA extraction kit.DNA concentration and purity were measured by UV spectrophotometry,DNA integrity detected by 1.25% agarose gel electrophoresis(OD260 / OD280 between 1.7 and 2.0,total > 1 ?g,concentration >50 ng/?L,If the DNA band is clear and there are no diffuse bands,it is qualified DNA).Specimens were stored in a refrigerator at-80 °C for later use.3.Bisulfite treatment: DNA samples were processed using the EpiTect Bisulfite Kit to modify DNA CpG islands.The total amount of DNA treatment per sample is 1 ?g,which can satisfy up to 200 candidate region fragment sequencing detection.4.Whole genome amplification,hybridization,scanning: The whole genome of each sample modified with sodium bisulfite was amplified,fragmented and digested;The treated samples were purified,precipitated,and resuspended using a PCR purification kit.5.Whole genome DNA methylation site screening: Hybridization,washing,extension,staining,and scanning were performed on an Illumina Infinium Methylation EPIC BeadChip to obtain DNA methylation signals.Application of cluster analysis,GO analysis,KEGG analysis for signal acquisition and chip results analysis.The DNA methylation ratio background values were corrected using a standard control site built into the Ilumina Infinium Methylation EPIC BeadChip.Results There were 67,984 CpG loci in the case group and the skin tissue control group,which differed in the degree of methylation.Among them,there were 30,621 CpG loci with increased methylation levels and 37,363 CpG loci with decreased methylation levels.The number of genes corresponding to the differential methylation loci was 4,635,while the number of genes with higher methylation level was 2,284,and the number of genes with lower methylation level was 2,351.The GO analysis lists the top 10 functional categories of differentially methylated genes.The KEGG pathway analysis showed that differentially methylated genes were mainly enriched in signaling pathways such as inflammation,sex hormones,and cancer.Conclusions There are DNA aberrant methylation-specific genes in skin tissue of AV patients.DNA aberrant methylation may be one of the important causes of AV pathogenesis,and it is the main way of epigenetic regulation of acne;In this study,Illumina Infinium Methylation EPIC BeadChip chip technology was used to preliminarily establish the genomic methylation genetic map of acne vulgaris skin tissue,and have a preliminary understanding of the function and signaling pathway of these genes.Laid the foundation for epigenetic pathogenesis,treatment and prevention of acne vulgaris.