骨肉瘤患者血清蛋白质组学的研究

Detection of serum biomakers of osteosarcoma by proteomic profiling

目的从蛋白质水平寻找和鉴定骨肉瘤患者血清中的生物学标志物，并初步建立骨肉瘤的血清蛋白质谱。方法采集8例血清蛋白样本（骨肉瘤患者4例及性别、年龄匹配的正常人对照组4例），用不同的CyDye荧光染料交叉标记后，依次进行双向胶内差异凝胶电泳（2-DDIGE）、图像分析和基质辅助激光解析离子化飞行时间质谱（MALDI-TOF-MS）鉴定。对质谱成功鉴定出的蛋白质，采用Westernblot检测方法，进一步验证有意义的差异性蛋白质。结果对2-DDIGE分析结果进行质谱鉴定，MALDI．TOF—MS成功鉴定出58个蛋白质，经MASCOT等数据库检索证实43个，其中上调18个，下调25个。Westernblot检测证实，凝胶溶素（gelsolin）在骨肉瘤血清中低表达。结论初步寻找到骨肉瘤患者血清中有意义的差异蛋白；gelsolin可能成为骨肉瘤的一个新的血清学诊断标志物，用于患者的大规模筛查，并为将来的基因治疗标定治疗靶点。

Objective To detect differentially expressed proteins in serum of patient vnth osteosarcoma. Methods 8 serum protein samples were recruited （4 cases of osteosarcoma and 4 cases of normal adults）, cross-labeled with variant CyDye, followed by two-dimensional differential in-gel electrophoresis （2-D DIGE ） , image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （ MALDI-TOF-MS ）. Results 24 protein spot-features were significantly increased, and 34 were significantly decreased in the serum from patients with osteosarcoma relative to the controls. The mass spectrometry analysis revealed 18 unique proteins that were increased, and 25 unique proteins decreased in the serum of patients with osteosarcoma. Gelsolin was down-regulated in osteosarcoma, and Western blotting also confirmed a decreased level of gelsolin in the serum of patients with osteosarcoma. Conclusion Our results indicate that gelsolin may have great potential as a biomarker of osteosarcoma and as a potential target for therapy. These preliminary data suggest that incorporation of more samples and new datasets will permit the identification of serum biomarkers for osteosarcoma.