【摘要】：目的通過(guò)檢測(cè)正常人、胃上皮內(nèi)瘤變、胃癌和胃癌術(shù)后患者血漿中聯(lián)合多基因的DNA甲基化水平,研究ZIC1,HOXD10和RUNX3在胃上皮細(xì)胞癌變過(guò)程中的基因甲基化動(dòng)態(tài)變化,探究血液基因甲基化作為一種無(wú)創(chuàng)分子標(biāo)記物在胃癌前病變和胃癌診斷及癌變監(jiān)測(cè)中的價(jià)值。方法收集正常健康人群、胃上皮內(nèi)瘤變、胃癌和胃癌術(shù)后患者血漿樣本,記錄患者年齡、性別、幽門(mén)螺桿菌(Hp)感染、病理分期分型等臨床病理資料。用甲基化特異性PCR(MSP)檢測(cè)ZIC1,HOXD10和RUNX3在血漿中的甲基化陽(yáng)性率,分析外周血DNA甲基化水平在胃癌發(fā)生發(fā)展中不同階段的變化情況,使用Fisher和Logistic回歸方法統(tǒng)計(jì)分析ZICl,HOXD10和RUNX3甲基化水平與患者臨床病理特征的相關(guān)性,利用ROC曲線分析對(duì)疾病診斷和預(yù)后判斷的價(jià)值。結(jié)果(1)共納入234例研究對(duì)象,包括正常的健康對(duì)照38例,平均年齡47.18±15.12歲;胃上皮內(nèi)瘤變患者56例,平均年齡59.71±10.62歲;胃癌臨床患者123例(40例早期胃癌和83例進(jìn)展期胃癌),平均年齡63.7±9.38歲;胃癌術(shù)后患者17例,平均年齡59.75±9.75。各組間年齡及性別構(gòu)成無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。(2)ZIC1、HOXD10和RUNX3在上皮內(nèi)瘤變組外周血中的啟動(dòng)子甲基化陽(yáng)性率分別是51.79%、33.92%、37.50%;在早期胃癌組中的陽(yáng)性率分別為67.50%、30.00%和54.21%;在進(jìn)展期胃癌組中的陽(yáng)性率分別為72.29%、54.21%和40.96%;正常對(duì)照組中上述基因均未見(jiàn)陽(yáng)性甲基化現(xiàn)象。胃癌患者ZIC1的甲基化水平較胃上皮內(nèi)瘤變組中有顯著升高(P0.05),HOXD10甲基化在早期和進(jìn)展期胃癌之間具有統(tǒng)計(jì)學(xué)差異(P0.05),且HOXD10在術(shù)后患者中外周血甲基化陽(yáng)性率為17.65%,顯著低于胃癌組中的陽(yáng)性率(P＜0.05)。(3)胃癌患者外周血漿中ZIC1、HOXD10和RUNX3的甲基化陽(yáng)性與否和患者的性別、年齡、腫瘤部位、腫瘤大小、分化程度、淋巴結(jié)轉(zhuǎn)移情況、TNM分期、CEA水平和Hp感染情況之間無(wú)顯著性關(guān)聯(lián)(P0.05)。其中僅HOXD10的甲基化陽(yáng)性與否同腫瘤的浸潤(rùn)深度呈正相關(guān)(OR=0.2518;95%CI:1.123-5.645,P=0.023)。(4)聯(lián)合檢測(cè)外周血漿中ZIC1、HOXD10和RUNX3的甲基化水平可以提高胃病變(胃癌+癌前病變)診斷的敏感性,三者聯(lián)合最高可達(dá)89.4%。在胃癌患者中,ZIC1 檢測(cè)的敏感性為 70.7%,高于 HOXD10(46.3%)和 RUNX3(42.3%),HOXD10 的診斷特異性(79.8%)高于 ZIC1(69.1%)和 RUNX3(78.7%)。聯(lián)合檢測(cè)ZIC1和HOXD10的診斷敏感性為83.1%,特異性為58.5%,AUC為0.703,對(duì)胃癌的診斷價(jià)值要均高于任一單獨(dú)指標(biāo)或其他聯(lián)合方式檢測(cè)。結(jié)論聯(lián)合檢測(cè)外周血漿中ZICl、HOXD10和RUNX3啟動(dòng)子的甲基化水平有望成為胃癌診斷的特異性方法,有助于胃癌高危人群的篩查、早期胃癌的診斷、胃癌癌變過(guò)程的監(jiān)測(cè)及術(shù)后療效的判斷。
[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 鹵15.12 years, 56 patients with gastric intraepithelial neoplasia with an average age of 59.71 鹵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 鹵9.38 years old, and 17 cases of postoperative gastric cancer patients with an average age of 59.75 鹵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.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R735.2
，

本文編號(hào)：2387554