CAJ | 학술논문

目的：应用质谱技术构建高发区食管癌及癌前病变血清蛋白质指纹图谱模型，为食管癌早期诊断寻找生物指标，并探讨差异蛋白在食管癌发生发展中的作用及意义。方法收集食管癌、癌前病变血清样本共254例，根据病理分为低级别上皮内瘤变（LGIN）、高级别上皮内瘤变（HGIN）、进展期食管鳞癌（AEC）、健康对照组。采用弱阳离子交换纳米磁珠（WCX）对血清蛋白进行纯化，MALDI-TOF-MS质谱进行检测，构建诊断模型并进行盲法验证。结果三组病变组与对照组以及各组间比较有统计学差异蛋白峰146个，由14个蛋白峰建立食管癌及癌前病变血清蛋白指纹图谱模型，蛋白峰分别是32721、3403、1772、1071、1414、33312、32023、31043、4662、2245、1307、10013、6868、1234。对早期食管癌及前病变的特异性为90.91%（30/33），对LGIN的灵敏度为100%（31/31），HGIN灵敏度为89.66%（52/58），AEC的灵敏度为81.26%（26/32），盲法预测验证结果表明；该模型对食管癌前病变及癌的特异性为95.652%（22/23），对LGIN的敏感性为95.455%（21/22），HGIN敏感性为94.118%（32/34），AEC的敏感性为95.238%（20/21）。经ExPasy数据库检索蛋白峰为32023，1234，1307，推测为ZAG（zinc alpha2 glycoprotein），Glycosyltransferase-like protein LARGE2，Fibrinogen alpha chain。结论采用MALDI联合WCX建立诊断模型具有高的灵敏度及特异性。根据各组间差异蛋白变化，推测食管癌的发生可能起始于LGIN或更早，提示对于表达与肿瘤相关差异蛋白峰的个体应给予定期随访。
목적：응용질보기술구건고발구식관암급암전병변혈청단백질지문도보모형，위식관암조기진단심조생물지표，병탐토차이단백재식관암발생발전중적작용급의의。방법수집식관암、암전병변혈청양본공254례，근거병리분위저급별상피내류변（LGIN）、고급별상피내류변（HGIN）、진전기식관린암（AEC）、건강대조조。채용약양리자교환납미자주（WCX）대혈청단백진행순화，MALDI-TOF-MS질보진행검측，구건진단모형병진행맹법험증。결과삼조병변조여대조조이급각조간비교유통계학차이단백봉146개，유14개단백봉건립식관암급암전병변혈청단백지문도보모형，단백봉분별시32721、3403、1772、1071、1414、33312、32023、31043、4662、2245、1307、10013、6868、1234。대조기식관암급전병변적특이성위90.91%（30/33），대LGIN적령민도위100%（31/31），HGIN령민도위89.66%（52/58），AEC적령민도위81.26%（26/32），맹법예측험증결과표명；해모형대식관암전병변급암적특이성위95.652%（22/23），대LGIN적민감성위95.455%（21/22），HGIN민감성위94.118%（32/34），AEC적민감성위95.238%（20/21）。경ExPasy수거고검색단백봉위32023，1234，1307，추측위ZAG（zinc alpha2 glycoprotein），Glycosyltransferase-like protein LARGE2，Fibrinogen alpha chain。결론채용MALDI연합WCX건립진단모형구유고적령민도급특이성。근거각조간차이단백변화，추측식관암적발생가능기시우LGIN혹경조，제시대우표체여종류상관차이단백봉적개체응급여정기수방。
Objective This study was to screen protein biomarkers in serum of esophageal carcinoma and precancerous lesions subjects using Mass Spectrometry in high incidence area of China. Boosting decision tree was constructed to identify normal from EC and precancerous lesions, to screen significant biomarkers for early diagnosis and explore the function in progression of esophageal cancer. Methods We recruited 254 subjects, Including low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia (HGIN), advanced esophageal carcinoma (AEC) and normal control (NOR). All serum samples were pretreated with weak cation exchange (WCX) magnetic beads and tested by MALDI-TOF-MS, finally construction of boosting decision tree and blinded testing. Results There were 146 protein peaks were detected between the three parents groups compared with normal, the decision tree classification were constructed by 14 protein peaks, which contain 32721, 3403, 1772, 1071, 1414, 33312, 32023, 31043, 4662, 2245, 1307, 10013, 6868, 1234. This decision algorithm correctly classified 90.91%(30/33) of normal control, 100% (31/31) of LGIN, 89.66% (52/58) of HGIN, and 81.26% (26/32) of AEC.In the blinded testing, the decision algorithm correctly classified 95.652%(22/23) of normal control, 95.455%(21/22) of LGIN, 94.118%(32/34) of HGIN, and 95.238%(20/21) of AEC. The peaks of 32023, 1234 and 1307 were speculated zinc alpha2 glycoprotein, glycosyltransferase like protein LARGE2 fibrinogen alpha chain and fibrinogen alpha chain by ExPasy database retrieval, others were unknown. Conclusion The diagnostic pattern are established by differently protein peaks could accurately recognize the patients groups and normal controls application of MALDI-TOF MS with high sensitivity and specificity. We speculate that the occurrence of esophageal carcinoma began in the LGIN or much earlier, the patients that have cancer-related proteins in serum should be follow up.