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
王景1,2,谢艳1,2,李培龙1,2,杜鲁涛1,2,王传新1,2
WANG Jing1,2, XIE Yan1,2, LI Peilong1,2, DU Lutao1,2, WANG Chuanxin1,2
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。
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