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目的:应用蛋白质组学双向凝胶电泳和质谱技术筛选前列腺癌骨转移血清标志物。方法:收集前列腺癌伴骨转移、不伴骨转移各5例血清样本。血清样品用除白蛋白试剂盒除去血清中的白蛋白后进行双向凝胶电泳,ImageMaster 2D Platinum软件分析,有意义的差异蛋白质点行基质辅助激光解析离子化飞行时间质谱(MALDI-TOF-MS)鉴定。结果:双向凝胶电泳显示,前列腺癌骨转移组血清蛋白中有15个斑点与前列腺癌无骨转移组有显著差异,前列腺癌骨转移患者血清中10个蛋白质表达水平显著增加,5个蛋白质表达水平显著下降。5个差异蛋白点进行胶内原位酶解,肽质量指纹图谱分析成功得到了3个蛋白质肽质量指纹图谱,并查询数据库初步鉴定了该3个蛋白质分别为锌α2糖蛋白、结合珠蛋白和载脂蛋白CⅢ。结论:双向凝胶电泳结合质谱鉴定是血清差异蛋白质组学研究的可靠平台和有力工具。所鉴定出的蛋白质与前列腺癌骨转移的发生、发展有关,可能是前列腺癌骨转移潜在的血清标志物。
OBJECTIVE: To screen serum markers of bone metastasis of prostate cancer by proteomic two-dimensional gel electrophoresis and mass spectrometry. Methods: Five cases of prostate cancer with bone metastasis and no bone metastasis were collected. Serum samples were removed with albumin removal serum albumin two-dimensional gel electrophoresis, ImageMaster 2D Platinum software analysis, meaningful differences in protein spots matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) Identification. Results: Two-dimensional gel electrophoresis showed that there were 15 spots in the serum protein of prostate cancer bone metastasis group and 10 cases of prostate cancer without bone metastasis group were significantly different. The expression of 10 proteins in serum of patients with prostate cancer bone metastasis increased significantly, and 5 proteins The level dropped significantly. Five different protein spots were digested in situ by in-gel method. Three peptide mass fingerprinting (RFLP) fingerprints were successfully obtained from peptide mass fingerprinting (RFLP) analysis. The three proteins were identified as zinc α2 glycoprotein, Apolipoprotein C III. Conclusion: Two-dimensional gel electrophoresis combined with mass spectrometry identification is a reliable platform and a powerful tool for differential proteomics research. The identified proteins are related to the occurrence and development of bone metastasis of prostate cancer and may be potential serum markers of bone metastasis of prostate cancer.