正常乳腺与乳腺癌核基质蛋白的双相电泳—飞行时间质谱研究

Analysis of differentially expressed nuclear matrix proteins of breast cancer and matched normal breast tissues by 2-DE and MALDI-TOF-MS

目的:应用比较蛋白质组学方法分析正常乳腺与乳腺癌组织的差异表达核基质蛋白,为人乳腺癌发病机制及其早期诊断的研究提供理论依据.方法:应用双相电泳(2-DE)每组分离4例乳腺组织核基质蛋白;应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)获得差异蛋白点的肽质量指纹图谱(PMF);通过Mascot软件查询NCBI数据库鉴定蛋白质.结果:每张图谱约检测到900个蛋白质点,共发现27个差异表达蛋白,选择背景清晰、重复性及分辨率较好、蛋白表达差异明显的点进行质谱分析,结合表观分子质量及等电点,初步鉴定了12种蛋白质.结论:这些差异蛋白质部分有望成为乳腺癌诊断及治疗的分子标志物.

AIM： To investigate the molecular mechanism of breast cancer （BC） and to screen biomarkers for the early diagnosis of BC by analyzing differentially expressed nuclear matrix proteins between BCs and matched normal breast tissues using comparative proteomic methods. METHODS： Four patients suffering from BC were included in this study. The nuclear matrix proteins were separated by two dimensional gel electrophoresis （2- DE） and differentially expressed proteins in the protein profiling were analyzed and identified by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. Proteins were identified by consulting NCBI database with Mascot software. RESULTS ： About 900 protein spots were seen on each gel. Twenty-~even protein spots with clear background, perfect repeatability, resolution and obvious protein expression difference were selected for further MALDI-TOF-MS analysis, with combination with molecular weight and isoelectric point. Twelve proteins were identified by comparative proteome analysis. CONCLUSION： The proteins identified may serve as potential biomarkers for the early diagnosis and therapy of breast cancer.