cg20657709位点甲基化对肺腺癌早期诊断的初步探讨

doi:10.6040/j.issn.1671-7554.0.2022.1418

摘要/Abstract

摘要： 目的探讨cg20657709位点甲基化在肺腺癌早期诊断中的临床价值。方法通过对30例肺腺癌患者(肺腺癌组)和45例健康对照者(健康对照组)进行Illumina Infinium MethylationEPIC BeadChip检测,获得基于外周血单个核细胞(PBMCs)的肺腺癌全基因组甲基化图谱以及候选差异甲基化位点(DMP)。进一步采用焦磷酸测序,在35例肺腺癌患者和30例健康对照者中,对该DMP对肺腺癌的诊断价值进行验证。最后,基于多重目的区域甲基化富集测序,在50例肺原位腺癌和50例健康对照者中,评估该DMP对肺原位腺癌的诊断价值。结果 Illumina Infinium MethylationEPIC BeadChip检测结果显示,肺腺癌组具有基于PBMCs的特异性甲基化图谱,肺腺癌组和健康对照组甲基化水平差异显著。以∣Δβ∣≥0.06且校正P<0.05为标准,初步筛选出1 345个DMPs。经过多步骤筛选,确定cg20657709为候选位点。cg20657709位点甲基化水平在肺腺癌患者中高表达(Δβ=0.097,校正P=0.004)。焦磷酸测序显示,cg20657709位点能够明显区分肺腺癌患者与健康对照者(AUC=0.946,敏感性为91.4%,特异性为90.0%,截断值为66.91)。多重目的区域甲基化富集测序显示,cg20657709位点对肺原位腺癌具有较好的诊断价值(AUC=0.787,敏感性为86.0%,特异性为64.0%,截断值为0.746 5)。结论与健康对照者相比,cg20657709位点甲基化水平在肺腺癌患者中显著升高,对肺腺癌的早期诊断具有重要的临床价值。

关键词: 肺腺癌, 外周血单个核细胞, 甲基化, 早期诊断, 临床价值

Abstract: Objective To investigate the clinical value of cg20657709 site methylation in the early detection of lung adenocarcinoma(LUAD). Methods The genome-wide DNA methylation profiling of peripheral blood mononuclear cells(PBMCs)and candidate differentially methylated position(DMP)were obtained from 30 LUAD patients and 45 healthy controls using Illumina Infinium MethylationEPIC BeadChip detection. Furthermore, the candidate DMP was validated in 35 LUAD patients and 30 healthy controls by pyrosequencing. Finally, multiple target region methylation enrichment sequencing was performed to evaluate the diagnostic value of DMP in the detection of lung adenocarcinoma in situ(AIS)in a cohort of 50 lung AIS patients and 50 healthy controls. Results Illumina Infinium MethylationEPIC BeadChip detection results showed that LUAD patients had specific genome-wide DNA methylation profiling based on PBMCs. The methylation levels of LUAD patients and healthy controls were significantly different. Initially, 1,345 DMPs were screened with the standard of∣Δβ∣≥0.06 and adjusted P<0.05. After multi-step screening, cg20657709 was determined as the candidate site. The cg20657709 site methylation was significantly higher in LUAD patients than in healthy controls (Δβ=0.097, adjusted P=0.004). Pyrosequencing results showed that cg20657709 site could clearly discriminate LUAD patients from healthy controls(AUC=0.946, sensitivity=91.4%, specificity=90.0%, cut-off value=66.91). Multiple target region methylation enrichment sequencing results showed cg20657709 site had good value in the detection of lung AIS(AUC=0.787, sensitivity=86.0%, specificity=64.0%, cut-off value=0.746 5). Conclusion Compared with healthy controls, LUAD patients have significantly higher cg20657709 site methylation, which is an important marker in the early detection of LUAD.

Key words: Lung adenocarcinoma, Peripheral blood mononuclear cell, Methylation, Early diagnosis, Clinical value

中图分类号:

R734.2

刘士标,张淑君,李培龙,杜鲁涛,王传新. cg20657709位点甲基化对肺腺癌早期诊断的初步探讨[J]. 山东大学学报 (医学版), 2023, 61(4): 18-25.

LIU Shibiao, ZHANG Shujun, LI Peilong, DU Lutao, WANG Chuanxin. The cg20657709 site methylation in the early detection of lung adenocarcinoma[J]. Journal of Shandong University (Health Sciences), 2023, 61(4): 18-25.

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