胃癌患者胃液中特异性荧光物质的分离鉴定  

Isolation and Identification of Specific Fluorescent Substances from Gastric Juice of Patients with Gastric Carcinoma

作　　者：张月宁[1] 金新莲[2] 孟灵梅[2] 耿秋明[3] 彭佳柔[4] 周丽雅[2] 林三仁[2] 

机构地区：[1]首都医科大学附属北京佑安医院消化科,北京市100069 [2]北京大学第三医院消化科 [3]北京大学第三医院中心实验室 [4]北京大学医学部蛋白质组学国家重点实验室

出　　处：《中国肿瘤临床》2011年第1期10-14,共5页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金资助(编号:30371603;30672395)~~

摘　　要：目的:探索胃癌患者胃液中特异性荧光物质的性质,以期初步了解胃癌特征性荧光的来源和机制。方法:选取2006年8月至2008年3月在我院胃镜检查的患者收集胃液。采用Mono Q离子交换联合C18反相高效液相色谱分析方法分离胃液中的肿瘤特异性荧光物质,进行基质辅助解吸附电离飞行时间以及正交加速电喷雾串联质谱分析,测定其所含物质的质荷比并进一步鉴定其一级肽段序列。结果:共收集病例118例,其中胃癌组56例(包括进展期胃癌49例,早期胃癌7例),对照组62例。Mono Q离子交换高效液相色谱条件下,胃液固有荧光光谱共出现4个主要荧光峰,第一峰位于保留时间14～21min之间,其荧光强度均值明显高于对照组(P<0.05);进一步进行C18-反相高效液相色谱分离,荧光光谱主要呈锯齿状的荧光峰群,在30～32min之间有一个强度最大的荧光峰,胃癌组荧光强度明显高于胃良性病变组(P<0.01)。收集对应保留时间的样品进行基质辅助激光解吸附电离-飞行时间质谱分析,结果显示,高效液相色谱法分离胃液后可得到质荷比在500～5 300之间的多种物质。电喷雾电离-串联质谱得到了其中13种目前未知的多肽序列。结论:造成胃良恶性疾病间产生差异荧光现象的机制复杂,一些含荧光基团的多肽类物质可能在其中发挥一定的作用。Objective： To explore the characteristics of the cancer-specific fluorescent substances found in the gastric juice of gastric cancer patients and to gain a preliminary understanding of the mechanisms involved in the gastric carcino-maspecific autofluorescence spectrum. Methods： Clinical and pathological data of patients with different gastric diseases undergoing gastroscopy in our hospital between August 2006 and March 2008 were collected. Gastric juice was also collected. Mono Q ion-exchange combined with C18 reversed-phase high performance liquid chromatography was used to isolate the cancer-specific substances from gastric juice. Substances obtained were analyzed by matrix-assisted taser-desorption ionization-flight time mass spectrometry and electrospray ionization mass spectrometry/mass spectrometry to identify their mass-to-charge ratio and peptide sequence. Results： One hundred and eighteen cases were enrolled in this study, of which 56 were gastric carcinoma cases （including 49 with advanced gastric cancer and 7 with early-stage cancer） and 62 were controls. Under the conditions of Mono Q ion-exchange high performance liquid chromatography, 4 fluorescent peaks were shown in the primary gastric juice fluorescence spectrum, in which the first one locating at the retention time of 14-21min was significantly stronger in the cancer groups than in the control groups （ P 〈 0.05 ）. The C18 reversed-phase high performance liquid chromatography presented a serrated fluorescence peak group in the fluorescent spectrum. The in- tensity of the fluorescent peak locating at the retention time of 30-32min was significantly stronger in the cancer groups than in the control groups. Matrix-assisted laser-desorption ionization flight time mass spectrometry revealed that the mass-to-charge ratio of these substances from gastric juice after the separation by high performance liquid chromatogra- phy was at a range between 500 and 5300. Sequences of 13 polypeptides currently unknown were obtained through elec- tro

关 键 词：胃癌 胃液 高效液相色谱 质谱 多肽 

分 类 号：R735.2[医药卫生—肿瘤]