Role Of DNA Methylation Modification Of PI3K-Akt Pathway Gene In Psoriasis

Objective To investigate the effect of DNA methylation of PI3K-Akt signal pathway genes on psoriasis,the methylation levels of genes with different DNA methylation in PI3K-Akt signal pathway and their CpG sites were examined,further screening of the possible pathogenic genes and CpG loci will pave the way for exploring new detection methods,therapeutic targets and predictive biomarkers of psoriasis.Methods Firstly,the differentially methylated genes detected by the 850 K chip were analyzed by the software cluster Profiler,and the differentially methylated genes were screened,the CpG sites(p < 0.05)of significantly different genes in the pathway were selected as candidate verification sites.The lesional skin(PP)and the adjacent normal skin(PC)of 30 patients with psoriasis were collected as verification samples,quality control of DNA samples was performed by agarose gel electrophoresis,photometer detection of DNA concentration and purity,and DNA samples were transformed by sulfite treatment.The primers designed based on“Primer3” software were optimized,and the transformed genome was used as template to perform multi-panel PCR reaction.The Index sequence was introduced to the end of the library by PCR amplification,and all samples were equally mixed to obtain the sequenced library.The sequencing data were processed by FASTQC software,and the methylation level data of candidate sites were obtained by filtering and comparing with Flash and Blat software,screening for differentially methylated sites.Results1.Pi3k-akt,Cakium,Ras and other signaling pathways were significantly enriched in the methylation levels of PP Group and PC Group,and the PI3K-AKT signaling pathway was the most enriched gene,the difference was the most significant;2.The Methyl Target technique was used to verify the reliable methylation sequencing data.The analysis of the methylation level difference of CpG sites showed that compared with the PC Group,the PP group had a higher methylation level than the PC group,the methylation levels of 30 CpG sites including additional sites were significantly different,17 of which were up-regulated and13 were down-regulated;3.The difference analysis of gene methylation level showed that there were 11 CpG loci with significant differences between PP group and PC group,among which 9 CpG loci were consistent with the previous genome-wide DNA methylation detection results.In the PP group,there are 5 up-regulated,namely cg15381320,cg24818418,cg17940858,cg25409939 and cg24116498.In the PP group,there are 4 down-regulated,namely cg09931511,cg14815878,cg24421822 and cg10219562;4.The methylation level of CpG loci of JAK1,EGFR,EGF,FGFR2,HGF,NFKB1,PRL,TLR4 and other genes changed;5.The richness of each methylated haplotype in CpG locus of PP group and PC group was tested for significance,and 34 haplotypes with significant statistical differences were detected.Conclusion1.DNA methylation of 1.PI3K-Akt pathway gene may play an important role in the occurrence and development of psoriasis;2.Nine CpG loci including cg15381320,cg24818418,cg17940858,cg25409939,cg24116498,cg09931511,cg14815878,cg24421822 and cg10219562 may become new therapeutic targets or biomarkers for psoriasis;3.JAK1,EGFR,EGF,FGFR2 HGF,NFKB1,PRL,TLR4 and other genes have the potential as therapeutic targets or biomarkers;4.The 34 methylated haplotypes detected may be related to the increased risk of psoriasis,and may also provide theoretical basis for finding DNA methylation detection reagents for psoriasis and finding new detection methods.