血浆cfDNA甲基化联合检测在肝癌诊断中的价值  

Evaluation of Diagnostic Performance of Plasma cfDNA Methylation Combined Detection in Diagnosis of Hepatocellular Carcinoma

作　　者：夏娜梅 赵言腾 杨乾坤[2] 董兰兰 张玮 黄月 臧文巧[1] XIA Namei;ZHAO Yanteng;YANG Qiankun;DONG Lanlan;ZHANG Wei;HUANG Yue;ZANG Wenqiao(School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450000,China;Department of Transfusion,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Research and Development Department,Wuhan Ammunition Life-Tech Co.,Ltd.,Wuhan 430000,China)

机构地区：[1]郑州大学基础医学院,河南郑州450000 [2]郑州大学第一附属医院输血科,河南郑州450000 [3]武汉艾米森生命科技有限公司研发部,湖北武汉430000

出　　处：《河南医学研究》2024年第11期1921-1925,共5页Henan Medical Research

基　　金：河南省科技攻关项目(232102310028)。

摘　　要：目的 探讨血浆循环游离DNA(cfDNA)中GNB4、TCF24、Riplet、ACP1和TSPYL5基因甲基化对肝癌诊断的临床价值。方法 利用滑动窗口技术在公共数据库中筛选肝癌和正常组织间的差异甲基化标志物并分析其甲基化水平。从郑州大学第一附属医院收集肝癌、肝硬化组织和健康人白细胞样本,Sanger测序检测筛选出的标志物的甲基化水平,选出敏感度和特异度较好的标志物。收集24例肝癌、23例肝硬化和99例健康人血浆,通过甲基化特异性PCR检测上述标志物单独和组合时对肝癌的诊断性能。结果 5个标志物(GNB4、TCF24、Riplet、ACP1和TSPYL5)在肝癌样本中显著高甲基化。组织测序结果显示除TCF24外,其他标志物在肝硬化组织中的甲基化阴性率均大于80.0%。在血浆样本验证中,GNB4单独诊断肝癌的曲线下面积(AUC)最大,为0.765,敏感度为66.7%,特异度为91.8%。在多基因组合中,GNB4+TSPYL5的诊断性能最佳,AUC值为0.893,敏感度为83.3%,特异度为90.2%。结论 GNB4、Riplet、ACP1和TSPYL5甲基化可用于肝癌诊断,且联合诊断性能优于单一基因。Objective To investigate the clinical value of methylation of GNB4,TCF24,Riplet,ACP1,and TSPYL5 genes in plasma circulating free DNA(cfDNA)for the diagnosis of liver cancer.Methods Sliding window technology was used to screen differential methylation markers between liver cancer and normal tissues in public databases and analyze their methylation levels.Liver cancer,liver cirrhosis tissue,and healthy human leukocyte samples were collected from the First Affiliated Hospital of Zhengzhou University,and Sanger sequencing was used to detect the methylation levels of selected biomarkers,biomarkers with better sensitivity and specificity were selected.Plasma from 24 cases of liver cancer,23 cases of liver cirrhosis,and 99 healthy individuals were collected,and the diagnostic performance of the above markers alone and in combination for liver cancer were detected through methylation specific PCR.Results Five biomarkers(GNB4,TCF24,Riplet,ACP1 and TSPYL5)were significantly hypermethylated in liver cancer samples.The tissue sequencing results showed that except for TCF24,the methylation negative rate of other biomarkers in liver cirrhosis tissue was greater than 80.0%.In plasma sample validation,GNB4 alone had the highest AUC for diagnosing liver cancer,at 0.765,with a sensitivity of 66.7%and a specificity of 91.8%.In the multi gene combination,GNB4+TSPYL5 had the best diagnostic performance,with an area under curve(AUC)value of 0.893,sensitivity of 83.3%,and specificity of 90.2%.Conclusion Methylation of GNB4,Riplet,ACP1 and TSPYL5 can be used for liver cancer diagnosis,and the combined diagnostic performance is better than that of a single gene.

关 键 词：肝癌 循环游离DNA 甲基化 血浆诊断 

分 类 号：R735.7[医药卫生—肿瘤]