DNA甲基化和胃液固有荧光光谱联合检测在胃癌诊断中的意义  

The value of p16 and death-associated protein kinase methylation and autofluroscene spectrum analysis of gastric juice in diagnosing gastric carcinoma

作　　者：闫秀娥[1] 周丽雅[1] 林三仁[1] 

机构地区：[1]北京大学第三医院消化科,100083

出　　处：《中华消化杂志》2006年第11期726-730,共5页Chinese Journal of Digestion

摘　　要：目的探索在胃癌患者胃液、外周血清中检测基因甲基化的可行性,并结合胃液稀释固有荧光光谱评价二者在胃癌诊断中的作用。方法采用甲基化特异性PCR方法,检测50例胃癌患者的原发肿瘤组织、外周血清和胃液脱落细胞的死亡相关蛋白(DAP)激酶、p16基因启动子区域甲基化状态,并以胃良性溃疡和慢性浅表性胃炎各20例、慢性萎缩性胃炎30例作为对照,同时检测胃癌患者和对照者的胃液稀释固有荧光光谱。结果50例胃癌患者的肿瘤组织、外周血清和胃液脱落细胞中p16和DAP激酶基因甲基化阳性率分别为74．4%和68．1%、52．0%和58．0%、58．6%和76．0%,20例慢性浅表性胃炎患者中均未检测到基因异常甲基化；20例胃溃疡患者溃疡周边组织、胃液脱落细胞中p16和DAP激酶甲基化阳性率为10．0%和20．0%、5．0%和15．0%,在外周血清中未检测到异常甲基化；30例慢性萎缩性胃炎患者胃黏膜组织、外周血清、胃液脱落细胞p16和DAP激酶基因甲基化阳性率分别为10．0%和23．3%、3．3%和3．3%、3．3%和20．0%。胃癌患者胃液固有荧光光谱强度较对照者明显增强(P＜0．05)；以P_1FI≥111．8为分界点分析胃液固有荧光光谱诊断胃癌的敏感性和特异性分别为76．1%和78．6%。p16和DAP激酶基因甲基化和胃液固有荧光光谱结合后诊断胃癌的敏感性提高到95．6%和97．8%。结论胃癌患者外周血清及胃液脱落细胞中可检测到与原发肿瘤组织一致的基因异常甲基化,胃液固有荧光光谱和DNA甲基化联合对检测胃癌有良好的临床应用价值。Objective To explore the feasibility of detecting death-associated protein （DAP） kinase promoter hypermethylation in the tumor tissues and plasma of patients with gastric adenocarcinoma, and to evaluate the effect of DNA hypermethylation and autofluroscene spectrums of gastric juice on diagnosing gastric carcinoma. Methods Primary tumor tissues and plasma and gastric juice of 50 patients with gastric adenocarcinoma were collected. Gastric mucosa tissue, plasma and gastric juice of 20 patients with chronic superficial gastritis and 20 patients with benign gastric ulcer and 30 patients with chronic atrophic gastritis were collected as controls. After sodium-bisulfite treatment, extracted DNA was amplified for DAP kinase promoter hypermethylation by methylation-specific polymerase chain reaction. At the same time the gastric juice autofluroscene spectrums （the excitation wavelength was 288 nm, whereas the range of emission wavelength was 300-800 nm） were detected. Results Among the samples from 50 patients, p16 and DAP kinase hypermethylation were detected in 74.4 % and 68. 1% of tumor tissues, 52.0% and 58.0% of plasma, 58.6% and 76.0% of gastric juice. No hypermethylation was detected in samples of chronic superficial gastritis and serum of patients with gastric ulcer. Among the patients with gastric ulcer, p16 and DAP kinase hypermethylation were detected in 10. 0% and 20.0% of tissues, 5.0% and 15.0% of gastric juice. Among the samples from 30 patients with chronic atrophic gastritis, p16 and DAP kinase were detected in 10.0% and 23.3% of tissues, 3.3% and 3.3% of sera, 3. 3% and 20. 0% of gastric juice. The intensity of gastric juice autofluroscence spectrums of patients with gastric carcinoma was much higher than those in controls. The sensitivity of p16 and DAP kinase hypermethylation and autofluorescence spectrums together were 95.6% and 97.8%, respectively. Conelusions Promoter hyperrnethylation of the p16 and DAP kinase gene detected in plasma and gastric juice consists with that in primary tumor t

关 键 词：胃肿瘤 DNA甲基化 荧光 诊断 

分 类 号：R735.2[医药卫生—肿瘤]