同源异形盒基因D10启动子甲基化与胆管癌患者预后的关系  

Relationship between promoter methylation of homeobox D10 gene and clinical prognosis of patients with cholangiocarcinoma

作　　者：杨海霞[1,2] 米建强[3] 梁军[1,2] 邵秋菊[1,2] 齐宇红[1,2] 常浩[1,2] 王启明 李恩孝[4] YANG Haixia;MI Jianqiang;LIANG Jun;SHAO Qiuju;QI Yuhong;CHANG Hao;WANG Qiming;LI Enxiao(Department of Radiotherapy, Tangdu Hospital, Fourth Military Medical University, Xi＇an 710038, Shaanxi Province, China;Cancer Institute, Fourth Military Medical University, Xi＇an 710038, Shaanxi Province, China;Department of Pathology, F＇trst Affiliated Hospital of Henan Science and Technology University, Luoyang 471003, Henan Province, China;Department of Medical Oncolog, First Affilited Hospital of Xi＇ an Jiaotong University, Xi＇ an 710061, Shaanxi Province, China)

机构地区：[1]第四军医大学唐都医院放射治疗科,陕西西安710038 [2]第四军医大学肿瘤研究所,陕西西安710038 [3]河南科技大学第一附属医院病理科,河南洛阳471003 [4]西安交通大学第一附属医院肿瘤内科,陕西西安710061

出　　处：《肿瘤》2018年第5期469-475,512,共8页Tumor

摘　　要：目的:研究同源异形盒基因D10(homeobox D10,HOXD 10)在人胆管癌组织中的启动子甲基化状态,并探讨HOXD 10基因启动子甲基化与胆管癌患者预后的关系。方法:采用甲基化特异性PCR法检测87例人胆管癌组织与30例癌旁组织中HOXD 10基因启动子的甲基化情况,并进一步采用重亚硫酸盐测序法检测HOXD 10基因启动子中27个胞嘧啶鸟嘌呤二核苷酸(cytosinephosphateguanosine,CpG)位点的甲基化程度。然后,应用Spearman相关检验分析HOXD 10基因启动子甲基化与其蛋白表达的相关性,用cutpoint生存分析和Kaplan-Meier法检验HOXD 10基因启动子区CpG位点甲基化数目与胆管癌患者预后的关系。结果:胆管癌组织中HOXD 10基因启动子甲基化率明显高于癌旁组织(P<0.05),而且癌旁组织中HOXD 10启动子区域的CpG位点数目少于胆管癌组织。HOXD10蛋白表达和HOXD 10基因启动子甲基化在胆管癌中呈显著负相关性(r=-0.82,P<0.000 1)。HOXD 10基因启动子区有7个CpG位点甲基化是胆管癌患者生存分析的最佳"截点"(P<0.001),CpG位点数目为0～6个的胆管癌患者生存时间明显长于CpG位点数目≥7个的患者(P<0.01)。结论:基因启动子甲基化与HOXD10蛋白的表达失活密切相关,HOXD 10基因启动子区存在≥7个CpG位点甲基化可作为胆管癌患者预后不良的预测指标之一。Objective： To investigate the methylated status of homeobox D10（HOXD 10） gene promoter in human cholangiocarcinoma tissues,and to explore the relationship between the promoter methylation of HOXD 10 gene and the clinical prognosis of patients withcholangiocarcinoma.Methods： The methylation of HOXD 10 gene promoter in 87 human cholangiocarcinoma tissues and 30 para-cancerous tissues was determined by methylation-specific PCR.Additionally, the methylation status of 27 cytosine-phosphate-guanosine（CpG） loci in HOXD 10 gene promoter was detected by bisulfite genomic sequencing PCR. Then the correlation between the protein expression and the promoter methylation of HOXD 10 gene was assessed by Spearman correlation test. Furthermore, the association of the number of methylated CpG loci with the survival of cholangiocarcinoma patients was examined by cut-point survival analysis and Kaplan-Meier test.Results： The positive methylation rate of HOXD 10 gene promoter in cholangiocarcinoma tissues was significantly higher than that in para-cancerous tissues（P 〈0.05）. The number of methylated CpG loci in para-cancerous tissues was less than that in cholangiocarcinoma tissues. The expression of HOXD10 protein was negatively correlated with the methylation status of HOXD 10 gene promoter（r =-0.82, P 〈0.000 1）. Methylation of seven CpG loci in HOXD 10 gene promoter was the optimal cut-off value for the survival analysis of cholangiocarcinoma patients（P 〈0.001）. The survival time of cholangiocarcinoma patients with 0-6 methylated CpG loci of HOXD 10 gene promoter was significantly longer than that of the patients with more than 7 methylated CpG loci（P 〈0.01）.Conclusion： The methylation of gene promoter is closely correlated with the inactivation of HOXD10 protein expression, and the methylation of ≥7 CpG loci in HOXD 10 gene promoter may be a potential predictor of poor prognosis in the patients with cholangiocarcinoma.

关 键 词：胆管上皮癌 DNA甲基化 预后 同源异形盒基因D10 

分 类 号：R730.261[医药卫生—肿瘤]