联合检测血浆中ZIC1、HOXD10和RUNX3启动子甲基化水平对胃癌及癌前病变的诊断价值探究

发布时间：2018-12-20 06:15

【摘要】：目的通过检测正常人、胃上皮内瘤变、胃癌和胃癌术后患者血浆中联合多基因的DNA甲基化水平,研究ZIC1,HOXD10和RUNX3在胃上皮细胞癌变过程中的基因甲基化动态变化,探究血液基因甲基化作为一种无创分子标记物在胃癌前病变和胃癌诊断及癌变监测中的价值。方法收集正常健康人群、胃上皮内瘤变、胃癌和胃癌术后患者血浆样本,记录患者年龄、性别、幽门螺杆菌(Hp)感染、病理分期分型等临床病理资料。用甲基化特异性PCR(MSP)检测ZIC1,HOXD10和RUNX3在血浆中的甲基化阳性率,分析外周血DNA甲基化水平在胃癌发生发展中不同阶段的变化情况,使用Fisher和Logistic回归方法统计分析ZICl,HOXD10和RUNX3甲基化水平与患者临床病理特征的相关性,利用ROC曲线分析对疾病诊断和预后判断的价值。结果(1)共纳入234例研究对象,包括正常的健康对照38例,平均年龄47.18±15.12岁;胃上皮内瘤变患者56例,平均年龄59.71±10.62岁;胃癌临床患者123例(40例早期胃癌和83例进展期胃癌),平均年龄63.7±9.38岁;胃癌术后患者17例,平均年龄59.75±9.75。各组间年龄及性别构成无统计学差异(P0.05)。(2)ZIC1、HOXD10和RUNX3在上皮内瘤变组外周血中的启动子甲基化阳性率分别是51.79%、33.92%、37.50%;在早期胃癌组中的阳性率分别为67.50%、30.00%和54.21%;在进展期胃癌组中的阳性率分别为72.29%、54.21%和40.96%;正常对照组中上述基因均未见阳性甲基化现象。胃癌患者ZIC1的甲基化水平较胃上皮内瘤变组中有显著升高(P0.05),HOXD10甲基化在早期和进展期胃癌之间具有统计学差异(P0.05),且HOXD10在术后患者中外周血甲基化阳性率为17.65%,显著低于胃癌组中的阳性率(P＜0.05)。(3)胃癌患者外周血浆中ZIC1、HOXD10和RUNX3的甲基化阳性与否和患者的性别、年龄、肿瘤部位、肿瘤大小、分化程度、淋巴结转移情况、TNM分期、CEA水平和Hp感染情况之间无显著性关联(P0.05)。其中仅HOXD10的甲基化阳性与否同肿瘤的浸润深度呈正相关(OR=0.2518;95%CI:1.123-5.645,P=0.023)。(4)联合检测外周血浆中ZIC1、HOXD10和RUNX3的甲基化水平可以提高胃病变(胃癌+癌前病变)诊断的敏感性,三者联合最高可达89.4%。在胃癌患者中,ZIC1 检测的敏感性为 70.7%,高于 HOXD10(46.3%)和 RUNX3(42.3%),HOXD10 的诊断特异性(79.8%)高于 ZIC1(69.1%)和 RUNX3(78.7%)。联合检测ZIC1和HOXD10的诊断敏感性为83.1%,特异性为58.5%,AUC为0.703,对胃癌的诊断价值要均高于任一单独指标或其他联合方式检测。结论联合检测外周血浆中ZICl、HOXD10和RUNX3启动子的甲基化水平有望成为胃癌诊断的特异性方法,有助于胃癌高危人群的筛查、早期胃癌的诊断、胃癌癌变过程的监测及术后疗效的判断。
[Abstract]:Objective to study the dynamic changes of gene methylation of ZIC1,HOXD10 and RUNX3 during gastric epithelial cell carcinogenesis by detecting the DNA methylation level in plasma of normal subjects, gastric intraepithelial neoplasia, gastric cancer and gastric cancer. To explore the value of blood gene methylation as a noninvasive molecular marker in the diagnosis and monitoring of precancerous lesions and gastric cancer. Methods Plasma samples were collected from healthy population, gastric intraepithelial neoplasia, gastric cancer and gastric cancer after operation. The clinical and pathological data were recorded, such as age, sex, Helicobacter pylori (Hp) infection, pathological staging, and so on. The methylation positive rate of ZIC1,HOXD10 and RUNX3 in plasma was detected by methylation specific PCR (MSP). The changes of DNA methylation level in peripheral blood in different stages of gastric cancer were analyzed. ZICl, was statistically analyzed by Fisher and Logistic regression method. The correlation between the levels of HOXD10 and RUNX3 methylation and the clinicopathological characteristics of the patients. The value of ROC curve analysis in the diagnosis and prognosis of the disease was evaluated. Results (1) A total of 234 subjects were included, including 38 healthy controls with an average age of 47.18 卤15.12 years, 56 patients with gastric intraepithelial neoplasia with an average age of 59.71 卤10.62 years. 123 cases of gastric cancer (40 cases of early gastric cancer and 83 cases of advanced gastric cancer), the average age was 63.7 卤9.38 years old, and 17 cases of postoperative gastric cancer patients with an average age of 59.75 卤9.75 years. There was no significant difference in age and sex composition among groups (P0.05). (2). The positive rate of promoter methylation of ZIC1,HOXD10 and RUNX3 in peripheral blood of intraepithelial neoplasia group was 51.799.33.92% and 37.50%, respectively. The positive rates in early gastric cancer group were 67.50% and 54.21%, and in advanced gastric cancer group were 72.29% and 40.96%, respectively. No positive methylation of these genes was found in the normal control group. The methylation level of ZIC1 in gastric cancer patients was significantly higher than that in gastric intraepithelial neoplasia group (P0.05), and HOXD10 methylation was significantly different between early and advanced gastric cancer (P0.05). The positive rate of HOXD10 in peripheral blood methylation was 17.65, which was significantly lower than that in gastric cancer group (P < 0. 05). (3). The methylation of ZIC1,HOXD10 and RUNX3 in peripheral plasma of gastric cancer patients and the sex of patients were significantly lower than those in gastric cancer group (P < 0. 05). There was no significant correlation between age, tumor location, tumor size, differentiation, lymph node metastasis, TNM stage, CEA level and Hp infection (P0.05). Only the methylation of HOXD10 was positively correlated with the depth of tumor invasion (OR=0.2518;). 95 CI: 1.123-5.645P0. 023). (4) combined detection of ZIC1,HOXD10 and RUNX3 methylation in peripheral plasma can improve the sensitivity of gastric disease (gastric precancerous lesion). The highest combination of the three levels can reach 89.4. In gastric cancer patients, the sensitivity of ZIC1 was 70.7, which was higher than that of HOXD10 (46.3%) and RUNX3 (42.3%). The diagnostic specificity of HOXD10 (79.8%) was higher than that of ZIC1 (69.1%) and RUNX3 (78.7%). The diagnostic sensitivity and specificity of combined detection of ZIC1 and HOXD10 were 83.1 and 0.703 respectively. Conclusion the combined detection of methylation of ZICl,HOXD10 and RUNX3 promoters in peripheral plasma is expected to be a specific method for the diagnosis of gastric cancer, which is helpful to the screening of high risk population of gastric cancer and the diagnosis of early gastric cancer. Monitoring of carcinogenesis of gastric cancer and judgment of postoperative curative effect.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2
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本文编号：2387554