GPR133基因甲基化在早期胃癌淋巴结转移预测中的作用  

Role of GPR133 gene methylation in predicting lymph node metastasis in early gastric cancer

作　　者：张洁 赵颖慧 董雅琪 李娟[1] 李培龙 杜鲁涛 ZHANG Jie;ZHAO Yinghui;DONG Yaqi;LI Juan;LI Peilong;DU Lutao(Department of Clinical Laboratory,The Second Hospital of Shandong University,Jinan 250033,Shandong,China;Department of Clinical Laboratory,Qilu Hospital of Shandong University,Jinan 250012,Shandong,China)

机构地区：[1]山东大学第二医院检验医学中心,山东济南250033 [2]山东大学齐鲁医院检验医学中心,山东济南250012

出　　处：《山东大学学报(医学版)》2025年第3期76-84,共9页Journal of Shandong University:Health Sciences

基　　金：山东省重点研发计划(2021ZLGX02);山东省博士后创新项目(202201010)。

摘　　要：目的通过定量检测GPR133基因甲基化位点,探讨其在早期胃癌淋巴结转移(lymph node metastasis,LNM)预测中的临床价值。方法回顾性选取2023年1月至2024年5月山东大学第二医院消化内科收治的100例T1期胃癌患者为研究对象;采用MethylationEPIC芯片分析T1期胃癌LNM阳性和阴性患者的手术组织样本,评估全基因组甲基化状态并识别差异甲基化位点(differentially methylated positions,DMPs);基于基因富集和功能预测确定候选DMPs;选用20例LNM阳性及80例阴性患者的福尔马林固定石蜡包埋(formalin-fixed and parrfin-embedded,FFPE)切片通过焦磷酸测序验证候选DMPs的甲基化水平;采用受试者操作特征曲线下面积(area under curve,AUC)评估DMPs在早期胃癌LNM诊断中的临床价值。结果MethylationEPIC芯片分析显示,T1期胃癌LNM阳性样本具有特异性甲基化图谱;以|Δβ|≥0.1且P<0.01为筛选标准,共初步筛选出1794个DMPs;根据基因匹配、TCGA数据库分析及cAMP binding途径的富集结果,明确GPR133基因上的cg00633768为候选位点;焦磷酸测序结果显示,cg00633768位点甲基化水平能够显著区分LNM阳性和阴性样本(AUC=0.869,敏感性64.3%,特异性100%,截断值0.6189)。结论与LNM阴性患者相比,GPR133基因的cg00633768位点在LNM阳性患者中呈显著高甲基化状态,对预测早期胃癌LNM具有重要的临床价值。Objective To explore the clinical value of quantitatively detecting methylation sites in the GPR133 gene for predicting lymph node metastasis(LNM)in early gastric cancer.Methods This retrospective study included 100 patients with T1 stage gastric cancer treated at the Department of Gastroenterology,Second Hospital of Shandong University,from January 2023 to May 2024.The MethylationEPIC array was used to analyze the whole-genome methylation status of surgical tissue samples from LNM-positive and LNM-negative patients,so as to identify differentially methylated positions(DMPs).Candidate DMPs were determined based on gene enrichment analysis and functional prediction.Pyrosequencing was performed on FFPE sections from 20 LNM-positive and 80 LNM-negative patients to validate the methylation levels of candidate DMPs.The clinical value of DMPs in diagnosing early gastric cancer lymph node metastasis was assessed using the area under the receiver operating characteristic curve(AUC).Results MethylationEPIC array analysis revealed a specific methylation pattern in T1 stage gastric cancer LNM-positive samples.Using a threshold of|Δβ|≥0.1 and P<0.01,a total of 1,794 DMPs were initially identified.According to the results of gene matching and TCGA database analysis and cAMP binding pathway enrichment,cg00633768 on GPR133 gene was identified as a candidate site.Pyrosequencing results showed that cg00633768 methylation level could significantly distinguish between LNMS positive and negative samples(AUC=0.869,sensitivity=64.3%,specificity=100%,cut-off value=0.6189).Conclusion The methylation level of cg00633768 in the GPR133 gene is significantly higher in LNM-positive patients compared to LNM-negative patients.This site has substantial clinical value for predicting lymph node metastasis in early gastric cancer.

关 键 词：早期胃癌 淋巴结转移 甲基化 G蛋白偶联受体 GPR133 

分 类 号：R735.2[医药卫生—肿瘤]