Abstract
BackgroundOsteosarcoma (OS) is the most common primary malignant tumor of bone in children and adolescents. However, the knowledge in diagnostic modalities has progressed less. To identify new biomarkers for the early diagnosis of OS as well as for potential novel therapeutic candidates, we performed a sub-cellular comparative proteomic research.MethodsAn osteosarcoma cell line (MG-63) and human osteoblastic cells (hFOB1.19) were used as our comparative model. Plasma membrane (PM) was obtained by aqueous two-phase partition. Proteins were analyzed through iTRAQ-based quantitative differential LC/MS/MS. The location and function of differential proteins were analyzed through GO database. Protein-protein interaction was examined through String software. One of differentially expressed proteins was verified by immunohistochemistry.Results342 non-redundant proteins were identified, 68 of which were differentially expressed with 1.5-fold difference, with 25 up-regulated and 43 down-regulated. Among those differential proteins, 69% ware plasma membrane, which are related to the biological processes of binding, cell structure, signal transduction, cell adhesion, etc., and interaction with each other. One protein--CD151 located in net nodes was verified to be over-expressed in osteosarcoma tissue by immunohistochemistry.ConclusionIt is the first time to use plasma membrane proteomics for studying the OS membrane proteins according to our knowledge. We generated preliminary but comprehensive data about membrane protein of osteosarcoma. Among these, CD151 was further validated in patient samples, and this small molecule membrane might be a new target for OS research. The plasma membrane proteins identified in this study may provide new insight into osteosarcoma biology and potential diagnostic and therapeutic biomarkers.
Highlights
Osteosarcoma (OS) is the most common primary malignant tumor of bone in children and adolescents
For human osteoblastic cell line hFOB1.19, Protein Digest, isobaric tag for relative and absolute quantitation (iTRAQ) Labeling, and Strong Cation Exchange Fractionation iTRAQ labeling was done according to the kit protocol (Applied Biosystems Inc., Foster City, CA) and the previously reported by Jonghwa Jin [21,22]
Production and characterization of plasma membrane (PM) derived from cell lines the growth characteristic of the two cell lines-MG-63 and hFOB1.19 is different, the cells were cultured with similar nutrition
Summary
Osteosarcoma (OS) is the most common primary malignant tumor of bone in children and adolescents. To identify new biomarkers for the early diagnosis of OS as well as for potential novel therapeutic candidates, we performed a sub-cellular comparative proteomic research. Only a few papers have reported comparative proteome research in OS, including our previous data obtain by comparative proteomic analysis of patient sera [4,5,6,7,8]. In some of these researches, tissue and cell lines were used. It is important to systematically study the PM proteins involved in OS
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