摘要
目的：骨肉瘤是最常见的骨原发恶性肿瘤,常常早期发生肺转移,预后极差。本课题对人类骨肉瘤细胞系与正常成骨细胞系质膜进行比较蛋白质组学研究。方法：体外培养人骨肉瘤细胞系(MG-63)和人成骨细胞系(hFOB1.19),双水相法提取细胞质膜蛋白质,经iTRAQ试剂标记后,通过多维液相色谱-串联质谱(MDLC/MS/MS)定量分析质膜蛋白质。结果：鉴定出342种非冗余蛋白质,其中63种蛋白质(10种上调蛋白,53种下调蛋白)表达差异大于1.5倍。63种蛋白质中69%来自质膜,参与细胞内外物质转换,信息交流,细胞识别,代谢调节,免疫应答等生物过程且相互作用。通过生物信息学分析,确定结点蛋白CD151,并在临床病理切片免疫组化分析中验证其在骨肉瘤细胞质膜中的表达。结论：本研究首次应用质膜蛋白质组学方法对骨肉瘤细胞质膜进行研究；结点蛋白CD151在未来的研究中可能成为新的肿瘤标志物及治疗靶点；所鉴定出的差异质膜蛋白质可能为骨肉瘤研究提供新的切入点。
Background:Osteosarcoma (OS) is the most common primary malignant tumor of bone in children and adolescents. However, the knowledge in diagnostic modalities and treatment methods is limited. To identify new biomarkers that facilitate the early diagnosis of OS and that maybe develop novel therapeutic candidates, we performed a subcellular comparative proteomic research. Methods:OS cell (MG-63) and human primary cultured osteoblastic cell (hFOB1.19) were used as a model. Plasma membrane (PM) was obtained by aqueous two-phase partition. Proteins were analyzed through iTRAQ-based quantitative differential LC/MS/MS. Results:342 non-redundant proteins were identified.63 were differentially expressed with 1.5-fold difference, with 10 up-and 53 down-regulated. For the differential proteins,69% ware plasma membrane. They are related to the biological processes of binding, cell structure, signal transduction, cell adhesion, etc., and are interacted with each other. One protein-CD151 located in net nodes was verified by Immnohistochemistry in osteosarcoma tissue of patients. Conclusion:It is the first time to use plasma membrane proteomics to study the OS membrane proteins according to our knowledge. CD151 might be a new biomarker for the diagnosis of OS and probably become a novel therapeutic candidate through further research. The plasma membrane proteins identified in this study may provide new insights into osteosarcoma cancer biology and potential diagnostic and treatment biomarkers.
引文
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