应用双向差异凝胶电泳和质谱技术筛选胰腺癌血清早期诊断标志物

Research of screening early diagnostic biomarkers of pancreatic cancer using 2D-DIGE and MALDI-TOF-MS analysis

目的应用双向差异凝胶电泳和质谱技术对胰腺癌、胰腺良性肿瘤、慢性胰腺炎患者和正常人群外周血清进行分析，寻找胰腺癌潜在的特异性早期诊断标志物。方法应用双向差异凝胶电泳（2D—DIGE）技术分离2009年5月-2011年4月40例胰腺癌患者、10例良性肿瘤患者、10例慢性胰腺炎患者和40例正常对照组人群外周血清蛋白质，应用基质辅助激光解析电离飞行时间质谱（MALDI—TOF—MS）技术对其中差异显著的蛋白进行鉴定。结果成功鉴定出3个胰腺癌差异表达蛋白：血色素结合蛋白（Hpx），人纤维凝胶蛋白3（FCN3）和血清淀粉样蛋白P成分（SAP），该组蛋白在胰腺癌组中呈高表达，在胰腺良性肿瘤组、慢性胰腺炎组和正常对照组中呈低表达。各点胰腺癌组表达分别较正常组高表达1．57、1．99、1．63倍（P〈0．05）。结论应用2D—DIGE技术联合MALDF—TOF—MS技术筛选胰腺癌患者外周血清中的特异性生物标志物的方法具有较好的重复性和稳定性。检测鉴定的差异蛋白质Hpx、FCN3和SAP可能是潜在的胰腺癌早期诊断血清特异性标志物。

Objective A comparative proteomie method was utilized to analyze serum proteins among pancreatic cancer patients, pancreatic benign tumor group, chronic pancreatitis group and normal control group to discover a new potential specific early diagnostic marker. Methods Comparative analysis on the pancreatic peripheral blood protein profiling from 40 pancreatic cancer patients, 10 benign tumor patients, 10 chronic pancreatitis patients and 40 cancer-free controls from May 2009 to April 2011 was carried out by 2D differential gel electrophoresis （2D- DIGE） and differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results Three differentially expressed proteins, Hemopexin （ Hpx）, Ficolin 3 （ FCN3 ） and Serum amyloid P-component （SAP） was identified. Those proteins were higher ex- pression in pancreatic cancer group compared with benign tumor group, chronic pancreatitis group and normal con- trol group. Each point in pancreatic cancer expression were 1.57, 1.99, 1.63 times than normal control expres- sion, respectively （ P 〈 0. 05 ）. Conclusions In this study, the identified proteins, Hpx, FCN3 and SAP may be as potential specific early diagnostic markers of pancreatic carcinoma. 2D-DIGE and MALDI-TOF-MS technology in screening specific serum tumor markers of pancreatic cancer has a well repeatability and stability.