Gene Expression Analysis Of Genital Lichen Sclerosus In Boys Based On Transcriptome Sequencing

Objective: In this study,we used the second-generation sequencing technology to analyse the transcriptome of the foreskin of boys with lichen sclerosus and its adjacent normal foreskin,obtained significantly differentially expressed genes,enriched functions and pathways,screened the most likely candidate genes and pathways involved in the occurrence of lichen sclerosus,and revealed the possible mechanism of its occurrence and provide an important basis for further molecular mechanism research.Methods: Boys with lichen sclerosus who underwent circumcision in the Department of Pediatric Surgery,Sichuan Provincial People’s Hospital from January 2020 to December2021 were selected as the research objects,and the diseased prepuce tissues and the adjacent normal prepuce tissues were collected.The inclusion criteria included:(1)Children with lichen sclerosus confirmed by pathological examination;(2)Boys with redundant prepuce,phimosis,hypospadias,concealed penis and concealed penis.Exclusion criteria:(1)The external genitalia of the children with acute local inflammatory changes;(2)External genitalia combined with other skin diseases;(3)Children with severe underlying diseases could not tolerate the operation.Transcriptome sequencing technology was used to screen the most likely candidate genes and pathways involved in the occurrence of lichen sclerosus,combined with significantly differentially expressed genes,go function enrichment,KEGG pathway enrichment,protein interaction network analysis and the known function of genes themselves,and finally the expression of candidate genes was verified by quantitative reverse transcription PCR.Results: In this study,25 pairs of samples were collected,and the age of the patients ranged from 4.9 to 13.8 years(mean age 9.3 ± 2.3 years).Five pairs of samples were unable to complete the experimental study due to tissue RNA degradation,and the remaining 20 pairs were successfully sequenced.A total of 2108 differentially expressed genes were obtained,including 1417 up-regulated genes and 691 down-regulated genes.Go function enrichment analysis showed that 504 functions were significantly enriched in biological processes,17 functions were significantly enriched in cellular component,and 49 functions were remarkably enriched in molecular functions.Enrichment analysis of KEGG pathway showed that 23 pathways were significantly enriched.Combining With the function of gene itself and previous research results,we screened IFNG,TNF,ZAP70,JAK3,MMP1,CXCL13,TYRP1,FASLG and RAC2 to verify the m RNA level of candidate genes by q PCR,and found that the expression trend of these candidate genes was consistent with the transcriptome results.The expressions of IFNG,TNF,ZAP70,JAK3,MMP1,CXCL13,FASLG and RAC2 were significantly up-regulated in the preputial lesions of lichen sclerosus(P < 0.05),while the expression of TYRP1 was significantly down-regulated in normal foreskin group(P <0.05).Conclusion: In this study,we systematically explored the gene expression profile of lichen sclerosus in boys based on transcriptome sequencing technology,and the results suggested that there were significant changes in the expression levels of IFNG、TNF、ZAP70、JAK3、MMP1、CXCL13、TYRP1、FASLG and RAC2 genes in the lesions of lichen sclerosus.The biological functions and pathways of these genes suggest that these genes may be involved in the occurrence of atrophic lichen sclerosus.This study preliminarily screened some key genes which may be involved in the pathogenesis of lichen sclerosus atrophicus and provided clues for the subsequent function and further mechanism research.