人胰腺癌胰液蛋白质差异表达分析

The Expression of Proteins in the Pancreatic Juice of Patients with Pancreatic Cancer

目的:应用蛋白质组学方法分析人胰腺癌患者胰液蛋白质表达。方法:通过内镜下逆行胰胆管造影术(ERCP)中放置鼻胰管引流收集患者胰液,利用双向凝胶电泳,分离5例胰腺癌、6例慢性胰腺炎和3例胆总管结石等量混合胰液(200μg)的总蛋白质,用图像分析软件识别差异表达的蛋白质;应用质谱仪得到相应的肽质指纹图谱,搜索数据库鉴定部分差异蛋白质点。结果:1)经ERCP留置鼻胰管成功收集胰液标本,胰液量约为35～200ml,Brad-ford法测定蛋白质浓度为0.8～4.6μg/μl。同一胰液标本(200μg)的3块胶在蛋白质点位置上有较好的重复性;5例胰腺癌、6例慢性胰腺炎和3例胆总管结石等量混合胰液自身比较后可重复的蛋白质点均在75%以上。2)对图像分析软件识别的9个差异蛋白质点进行基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析,鉴定出胰腺癌相关的差异表达蛋白质。结论:人胰腺癌、慢性胰腺炎和胆总管结石患者胰液的蛋白质谱存在一定的差异。

Objective： To analyze the differential expression of proteins in the pancreatic juice of patients with pancreatic cancer （pc）, chronic pancreatitis （cp） and choledocholithiasis hy methods of proteomics. Methods： pancreatic juice was collected by naso-pancreatic drainage during endoscopic retrograde cholangiopancreatography. The total proteins of pooled pancreatic juice （200ug） with equal amount from five pc patients, six cp patients anti three chuledocholithiasis patients were separated by two-dimensional gel electrophoresis（2-DE） respectively. The expression of proteins were analyzed by image analysis software and identified by mass spectrometry and database searching. Results： 1） Pancreatic juice was successfully collected by naso-pancreatic drainage during endoscopic retrograde cholangiopancreatography with amount ranging from 35 to 200 ml and protein concentration ranging from 0.8-4.6μg/μl. Three gels of the same pancreatic juice （200ug） had good reproducibility in protein location. Rates of matched proteins from pooled pancreatic juice with equal amount from five pc patients, six cp patients and three choledocholithiasis patients all exceeded 75% by self-contrast. 2） Nine differential proteins obtained from image analysis software were analyzed by MALDI-TOF-MS anti some were identified as differential expressed proteins related to pancreatic cancer. Conclusion： Protein spectra of pancreatic juice from pc patients, cp patients and choledocholithiasis patients are different.