title: Plasma membrane proteomic analysis of human osteosarcoma and osteoblastic cells: revealing NDRG1 as a marker for osteosarcoma.
abstract: Osteosarcoma (OS) is the most common primary malignant bone tumor in children and adolescents. To identify new biomarkers for early diagnosis of OS and novel therapeutic candidates, we carried out a plasma membrane proteomic study based on two-dimensional electrophoresis (2DE). The OS cell line MG-63 and the human osteoblastic cell line hFOB1.19 were adopted as the comparison model. We extracted plasma membrane by aqueous two-phase partition extraction. The proteins were separated through 2DE. We analyzed the differentially expressed proteins by Imagemaster software and then identified them by liquid chromatography-tandem mass spectrometry, and the location and function of differential proteins were searched through the Gene Ontology database. In total, 220 protein spots were separated by 2DE. Seven proteins with more than 2.0-folds of difference were successfully identified from 13 gel spots, with 6 up-regulated and 1 down-regulated. Gene Ontology analysis of the differentially expressed proteins indicated that these proteins were involved in seven kinds of functions including binding, structural, cell motility, receptor activity, electron carrier activity, NADH dehydrogenase (ubiquinone) activity, and transcription repressor activity. The up-regulation of NDRG1 was verified in osteosarcoma through Western blotting and by immunohistochemistry in paraffin-embedded tissues. The plasma membrane proteins identified in this study may provide new insights into osteosarcoma cancer biology and potential diagnostic and therapeutic biomarkers.
authors.affiliation: Musculoskeletal Oncology Center, Shanghai 10th People's Hospital, Tongji University School of Medicine, Shanghai, 200072, China.
authors.forename: Yingqi
authors.initials: Y
authors.lastname: Hua
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Jia
authors.forename: Mengxiong
authors.initials: M
authors.lastname: Sun
authors.forename: Longpo
authors.initials: L
authors.lastname: Zheng
authors.forename: Lin
authors.initials: L
authors.lastname: Yin
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhengdong
authors.initials: Z
authors.lastname: Cai
mesh_terms: D000293:Adolescent; D000328:Adult; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000096:biosynthesis*; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002853:Chromatography, Liquid; D019295:Computational Biology / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D010006:Osteoblasts / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D015854:Up-Regulation; D055815:Young Adult; D000099259:N-myc Downstream-Regulated Gene 1 Protein
chemical_list: D014408:Biomarkers, Tumor; D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D000099259:N-myc Downstream-Regulated Gene 1 Protein