胰腺癌细胞内膜系统膜蛋白的蛋白质组学分析

Proteomic analysis of membrane proteins of endomembrane system in human pancreatic cancer cell

目的对胰腺癌细胞的内膜系统膜蛋白进行蛋白质组学鉴定分析。方法提取Capan-1、MiaPaCa-2、Panc-1三株胰腺癌细胞的膜蛋白，应用二维电泳（2-DE）技术将三株细胞的膜蛋白混合液分离，切取所获得的混合膜蛋白2-DE图上肉眼可见的蛋白位点，进行基质辅助激光解吸离子化飞行时间质谱鉴定、肽质指纹库（PMF）匹配，并在NCBI与ExPASy蛋白质文库中加以验证分析，获得其基因序列、蛋白序列、相对分子质量、等电点、细胞内定位、生物学功能等信息。结果在混合膜蛋白2-DE图上共有166个肉眼可见蛋白点，其中49个（29．5％，49／166）为ExPASy蛋白数据库中收录的可靠蛋白，分别为通道载体蛋白（4个）、跨膜信号转导通路蛋白（5个）、转录调节及翻译修饰蛋白（7个）、增殖凋亡通路蛋白（4个）、侵袭转移相关蛋白（2个）、细胞骨架蛋白（3个）、细胞代谢通路蛋白（14个）及功能不明蛋白（10个）。结论胰腺癌细胞的内膜系统膜蛋白广泛参与肿瘤细胞的代谢、增殖、变形、迁移、黏附等多种恶性生物学行为，可能成为候选的胰腺癌肿瘤标志物及治疗靶点。

Objective To analyze and identify the membrane proteins of endomembrane system in human pancreatic cancer cell by proteomies. Methods Membrane protein was extracted from pancreatic cancer cell lines Capan-1 ,MiaPaCa-2,Pane-1. The membrane protein mixture of the three pancreatic cancer cell lines were separated by two-dimensional electrophoresis（2-DE）. Positive dots of staining 2-DE gel were identified by MALDI-TOF mass spectrometry and PMF matching, and then evaluated by bio-informatics searching in NCBI and ExPASy databases. Information of membrane proteins were acquired like sequence, molecular weight,isoelectric point,location and biological functions. Results Forty-nine membrane proteins out of 166 protein dots which could be seen on the 2-DE gel were identified as channel carrier proteins （4 proteins）, signal transduetion proteins （5 proteins）, transcription regulatory and translation modification protein （7 proteins ）, proliferation and apoptosis related proteins （4 proteins ）, invasion and migration associated proteins （ 2 proteins ） , cytoskeleton proteins （ 3 proteins ） , metabolism pathway proteins （14 proteins）,and function unknown protein （10 proteins）. Conclusions Endomembrane proteins of pancreatic cancer cell play key roles in tumor malignant behavior like proliferation, metabolism, motility, adhesion and migration. These membrane proteins might become candidate biomarkers or targets of therapy of pancreatic cancer.