Comparative proteomic analysis of plasma membrane proteins between human osteosarcoma and normal osteoblastic cell lines.
BACKGROUND
Osteosarcoma (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.


METHODS
An 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.


RESULTS
342 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.


CONCLUSIONS
It 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.
Zhang
Zhiyu
Z
Department of Orthopaedics, The 4th Affiliated Hospital, China MedicalUniversity, Shenyang, 110032, China. davidzhang77@sohu.com
Zhang
Lijun
L
Hua
Yingqi
Y
Jia
Xiaofang
X
Li
Jian
J
Hu
Shuo
S
Peng
Xia
X
Yang
Pengyuan
P
Sun
Mengxiong
M
Ma
Fang
F
Cai
Zhengdong
Z
D015703:Antigens, CD / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001859:Bone Neoplasms / Q000378:metabolism*; D002411:Cation Exchange Resins; D045744:Cell Line, Tumor; D002852:Chromatography, Ion Exchange; D056148:Chromatography, Reverse-Phase; D019295:Computational Biology; D030562:Databases, Protein; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism*; D010449:Peptide Mapping; D010865:Pilot Projects; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D060185:Tetraspanin 24
D015703:Antigens, CD; D014408:Biomarkers, Tumor; C554715:CD151 protein, human; D002411:Cation Exchange Resins; D008565:Membrane Proteins; D060185:Tetraspanin 24
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