人乳头瘤病毒介导宫颈癌中甲基化修饰的诊断标志物的鉴定

Identifying diagnostic biomarkers of human papillomavirus mediated methylation modification in cervical cancer

目的:探讨人乳头瘤病毒(HPV)持续感染导致宫颈癌发展的分子变化及潜在的诊断标志物。方法:分别从GSE138080和GSE99511数据集中获取HPV阳性的宫颈上皮、高级别癌前病变(CIN)和鳞状细胞癌(SCC)的宫颈样本的基因表达谱和甲基化谱数据,鉴定在CIN和HPV阳性间及SCC和CIN间差异表达的基因和差异甲基化的位点。对差异表达基因进行共表达分析,筛选与疾病进展显著相关的模块,并对模块基因进行富集分析。进一步在模块中筛选可能受到甲基化修饰的基因,并绘制受试者工作特征(ROC)曲线。收集自2022年11月至2023年9月于本院接受治疗的30例患者的宫颈组织样本,包含10例HPV阳性的宫颈上皮、10例CIN 3级和10例SCC组织样本。利用实时定量聚合酶链反应(RT-qPCR)检测基因的表达。结果:共有1832个基因在CIN和HPV阳性之间及SCC和CIN之间同时差异表达。富集分析显示,D4S234E、DOCK9、KLK10、NUCKS1为可能受到甲基化修饰的CIN和HPV阳性之间差异表达基因,TM7SF2为可能受到甲基化修饰的SCC和CIN之间差异表达基因。ROC结果提示,D4S234E、DOCK9、KLK10、NUCKS1和TM7SF2对CIN具有良好的诊断作用。D4S234E、DOCK9、KLK10、TM7SF2在SCC中的表达最低,在HPV阳性中的表达最高(P<0.05);NUCKS1在SCC中的表达最高,在HPV阳性中的表达最低(P<0.05)。结论:D4S234E、DOCK9、KLK10、TM7SF2、NUCKS1可能为宫颈癌提供了潜在的早期诊断标志物。

Objective:To investigate the molecular alterations and potential biomarkers associated with the progression of cervical cancer due to persistent human papillomavirus(HPV)infection.Methods:Gene expression and methylation profiles were analyzed from normal HPV positive cervical epithelium,high-grade precancerous lesions(CIN),and squamous cell carcinoma(SCC)were obtained from the GSE138080 and GSE99511 datasets,respectively.Differentially expressed genes and differentially methylated probes were identified between CIN and HPV positive,as well as SCC and CIN samples.Co-expression analysis on differentially expressed genes was performed,modules significantly associated with disease progression were screened,and enrichment analysis was performed.Genes within these modules potentially subject to methylation were further analyzed,and receiver operating characteristic(ROC)curves were generated.From November 2022 to September 2023,30 cervical tissue samples were collected from our hospital,including 10 HPV positive cervical epithelium samples,10 CIN3 samples,and 10 SCC tissue samples.Real time-quantitative PCR(RT-qPCR)was used to detect gene expression.Results:A total of 1832 genes were differentially expressed between CIN and HPV positive,as well as between SCC and CIN simultaneously.Enrichment analysis indicated that D4S234E,DOCK9,KLK10,and NUCKS1 may be differentially expressed genes subject to methylation modifications between CIN and HPV positive,while TM7SF2 may be a differentially expressed gene subject to methylation modifications between SCC and CIN.The ROC results suggest that D4S234E,DOCK9,KLK10,NUCKS1,and TM7SF2 have good diagnostic value for CIN.D4S234E,DOCK9,KLK10 and TM7SF2 exhibit the lowest expression in SCC and the highest expression in HPV positive(P<0.05),whereas NUCKS1 shows the highest expression in SCC and the lowest expression in HPV positive(P<0.05).Conclusion:D4S234E,DOCK9,KLK10,TM7SF2,NUCKS1 may potentially serve as early diagnostic markers for cervical cancer.