CAJ | 학술논문

目的探讨内部萃取电喷雾电离质谱(iEESI-MS)分析技术在快速区分食管癌组织与癌旁正常组织边界中的应用价值,研究磷脂酰胆碱(POPC)等磷脂类物质差异性表达的生物学意义.方法采用自主设计的取样器和自主开发的iEESI-MS技术,在不进行样本预处理的前提下,对我院26例食管癌患者配对的食管癌组织及癌旁正常组织进行质谱分析;iEESI-MS参数:正离子模式,质谱扫描范围质荷比(m/z)为50～2 000 Da;电离电压为+4.5 kV;离子传输管温度为150℃;离子传输管电压为35 V;透镜电压为100 V;其他质谱参数均由LTQ-MS系统自动优化,获得其iEESIMS指纹谱图;用Matlab软件对样本iEESI-MS指纹谱图数据进行分析,得到主成分分析(PCA)结果;相对丰度数据用SPSS 19.0软件进行分析.结果本方法能在1min完成精确取样、质谱检测和数据分析;食管癌组织与癌旁正常组织在m/z 567、611、655、699、743、771、783、787、799、882等处的相对丰度差异有统计学意义(P＜0.05),其中食管癌组织中m/z 783、799比较于癌旁正常组织显著增加,多级碰撞诱导解离实验(CID)证实m/z 783为磷脂酰胆碱钠盐(POPC+ Na+),m/z 799为磷脂酰胆碱钾盐(POPC+ K+),食管癌组织中POPC+ Na+及POPC+K+在质谱指纹谱中相对丰度高于癌旁正常组织(m/z 783:7.45±1.65比0.82±0.21,P＜0.05;m/z 799:8.86±1.37比1.23±1.46),差异有统计学意义(P＜0.05);PCA区分两组样本的准确率达96.15％ (25/26).结论结合组织精密取样器和iEESI-MS技术,可快速得到稳定的食管癌及癌旁正常食管组织质谱信号,基于POPC等磷脂类物质差异表达可区分食管癌组织边界.
목적 탐토내부췌취전분무전리질보(iEESI-MS)분석기술재쾌속구분식관암조직여암방정상조직변계중적응용개치,연구린지선담감(POPC)등린지류물질차이성표체적생물학의의.방법 채용자주설계적취양기화자주개발적iEESI-MS기술,재불진행양본예처리적전제하,대아원26례식관암환자배대적식관암조직급암방정상조직진행질보분석;iEESI-MS삼수:정리자모식,질보소묘범위질하비(m/z)위50～2 000 Da;전리전압위+4.5 kV;리자전수관온도위150℃;리자전수관전압위35 V;투경전압위100 V;기타질보삼수균유LTQ-MS계통자동우화,획득기iEESIMS지문보도;용Matlab연건대양본iEESI-MS지문보도수거진행분석,득도주성분분석(PCA)결과;상대봉도수거용SPSS 19.0연건진행분석.결과 본방법능재1min완성정학취양、질보검측화수거분석;식관암조직여암방정상조직재m/z 567、611、655、699、743、771、783、787、799、882등처적상대봉도차이유통계학의의(P＜0.05),기중식관암조직중m/z 783、799비교우암방정상조직현저증가,다급팽당유도해리실험(CID)증실m/z 783위린지선담감납염(POPC+ Na+),m/z 799위린지선담감갑염(POPC+ K+),식관암조직중POPC+ Na+급POPC+K+재질보지문보중상대봉도고우암방정상조직(m/z 783:7.45±1.65비0.82±0.21,P＜0.05;m/z 799:8.86±1.37비1.23±1.46),차이유통계학의의(P＜0.05);PCA구분량조양본적준학솔체96.15％ (25/26).결론 결합조직정밀취양기화iEESI-MS기술,가쾌속득도은정적식관암급암방정상식관조직질보신호,기우POPC등린지류물질차이표체가구분식관암조직변계.
Objective To discuss the application of internal extractive electrospray ionization mass spectrometry (iEESI-MS) for distinguishing the esophageal cancer margins rapidly and study the difference of the phosphatidylcholine (POPC) between the esophageal cancerous and normal tissues.Methods The samples from 26 esophageal cancer patients were prepared for iEESI-MS.The iEESI-MS ion source mode was set to positive ion mode, mass rang was set to 50-2 000 Da, atomization voltage was set to +4.5 kV, ion transmission tube temperature was set to 150 ℃C, ion transmission tube voltage was set to 35.0 V, and lens voltage was set to 100.0 V.The data processing method of principal component analysis (PCA) were performed for the metabolic fingerprinting spectrmn.And the data of the relative abundance were processed by SPSS 19.0.Results This method could be completed in 1 min to sample accurately, mass spectrum detection and data analysis.The relative abundance of esophageal cancer tissue and normal tissue had obvious difference in the mass-to-change ratio (m/z) 567,611,655,699,743,771,783, 787, 799, and 882.Multi-stage collision induced dissociation (CID) experiment confirmed the m/z 783 was POPC + Na + and m/z 799 was POPC + K + , and the relative abundances of m/z 783 (POPC + Na + , 7.45 ± 1.65 vs.0.82 ± 0.21, P ＜ 0.05) and m/z 799 (POPC + K + , 8.86 ± 1.37 vs.1.23 ± 1.46, P ＜ 0.05) were greater in cancerous tissue.Moreover, two groups of samples of 25 patients were obviously different by PCA and the identification rate reached 96.15％.Conclusion Mass spectrum signal can be obtained rapidly from esophageal cancer and normal esophageal tissue by the technology of sampling accurately and iEESI-MS.We can distinguish the esophageal cancer margins by the difference relative abundances of POPC.