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山东大学学报 (医学版) ›› 2022, Vol. 60 ›› Issue (6): 26-34.doi: 10.6040/j.issn.1671-7554.0.2021.0110

• 临床医学 • 上一篇    下一篇

GZMB基因cg16212145位点的异常甲基化芯片测定对胃癌早筛的价值

王景1,2,谢艳1,2,李培龙1,2,杜鲁涛1,2,王传新1,2   

  • 发布日期:2022-06-17
  • 通讯作者: 王传新. E-mail:cxwang@sdu.edu.cn
  • 基金资助:
    山东省重点研发计划(2019GHZ003);山东大学临床研究项目(2020SDUCRCA002)

Value of abnormal methylation of cg16212145 on GZMB gene in early screening of gastric cancer

WANG Jing1,2, XIE Yan1,2, LI Peilong1,2, DU Lutao1,2, WANG Chuanxin1,2   

  1. 1. Department of Clinical Laboratory, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250033, Shandong, China;
    2. Tumor Marker Detection Engineering Laboratory of Shandong Province, Jinan 250033, Shandong, China
  • Published:2022-06-17

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摘要： 目的 获得外周血单个核细胞(PBMC)来源的胃癌特异性全基因组甲基化图谱,探讨PBMC来源的甲基化位点在胃癌早期诊断中的临床价值。 方法 采用850k甲基化芯片检测20例胃癌患者和20例健康对照者PBMC的全基因组甲基化状态,初步构建PBMC来源的胃癌特异性甲基化图谱,获得差异甲基化位点(DMPs)。对DMPs所在的基因进行基因本体(GO)分析及京都基因与基因组百科全书(KEGG)分析,进行功能预测。利用基于随机森林的多因素过滤方法对DMPs进行统计筛选。对34例Ⅰ期胃癌患者和21例健康对照者进行基于二代测序的多重目的区域甲基化富集测序,验证筛选得到的位点。 结果 甲基化芯片结果显示,与健康对照者比较,胃癌患者的PBMC具有独特的全基因组甲基化图谱,以|Δβ|>0.06、adjust P<0.05作为预筛选条件共得到5 883个DMPs,包括2 513个高甲基化位点和3 370个低甲基化位点;GO分析和KEGG分析显示,5 883个DMPs所在的2 677个基因主要与MAPK信号通路、癌症中的转录失调相关。颗粒酶B(GZMB)基因cg16212145位点在胃癌患者的PBMC中呈高甲基化水平,差异具有统计学意义(Δβ=0.080 7, adjust P=0.001)。多重目的区域甲基化富集测序结果表明,CpG位点cg16212145的异常甲基化能够区分Ⅰ期胃癌患者与健康对照者(AUC=0.807, P<0.001)。 结论 胃癌患者具有PBMC来源的特异性全基因组甲基化图谱,GZMB基因cg16212145位点在胃癌患者的PBMC中呈现高甲基化水平,在胃癌的早期筛查中具有重要临床价值。

关键词: 胃癌, DNA甲基化, 外周血单个核细胞, 早期筛查, 标志物

Abstract: Objective To obtain the specific genome-wide methylation profile of gastric cancer derived from peripheral blood mononuclear cells(PBMC), and to explore the clinical value of methylation points derived from PBMC in the early diagnosis of gastric cancer. Methods The genome-wide DNA methylation status of PBMC in 20 gastric cancer patients and 20 healthy controls was detected with Methylation850 BeadChip. The gastric cancer-specific methylation profile was constructed and differentially methylated points(DMPs)were obtained. Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed on the genes where DMPs were located for functional prediction. Statistical screening of DMPs was performed using a random forest-based multi-factor filtering method. Next-generation sequencing-based multiplex target region methylation enrichment sequencing was performed on 34 patients with stage I gastric cancer and 21 healthy controls to verify the selected DMPs. Results The results of methylation BeadChip showed PBMC from gastric cancer patients had unique genome-wide methylation profile. A total of 5 883 DMPs were obtained with |Δβ|>0.06 and adjust P<0.05 as pre-screening conditions, including 2 513 hypermethylated sites and 3 370 hypomethylated sites. GO analysis and KEGG analysis showed that 2 677 genes where 5 883 DMPs were located were mainly associated with MAPK signaling pathway and transcriptional dysregulation in cancer. Granzyme B(GZMB)gene cg16212145 was hypermethylated in PBMC of gastric cancer patients, and the difference was statistically significant(Δβ=0.0807, adjust P=0.001). The results of multiple target region methylation enrichment sequencing showed that abnormal methylation status of CpG site cg16212145 could distinguish patients with stage I gastric cancer from healthy controls(AUC=0.807, P<0.001). Conclusion Gastric cancer patients have a specific genome-wide methylation profile derived from PBMC. The GZMB gene cg16212145 exhibits high methylation status in PBMC of gastric cancer patients, which has important clinical value in the early screening of gastric cancer.

Key words: Gastric cancer, DNA methylation, Peripheral blood mononuclear cell, Early screening, Markers

中图分类号: 

  • R735.2
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