Comparative proteomic analysis of hypertrophic chondrocytes in osteoarthritis

被引:53
作者
Tsolis, Konstantinos C. [1 ,2 ]
Bei, Ekaterini S. [3 ]
Papathanasiou, Ioanna [4 ,5 ]
Kostopoulou, Fotini [4 ,5 ]
Gkretsi, Vassiliki [5 ]
Kalantzaki, Kalliopi [3 ]
Malizos, Konstantinos [6 ]
Zervakis, Michalis [3 ]
Tsezou, Aspasia [4 ,5 ]
Economou, Anastassios [1 ,2 ]
机构
[1] Inst Mol Biol & Biotechnol FoRTH, Iraklion, Greece
[2] KULeuven, Rega Inst Med Res, Dept Microbiol & Immunol, Leuven, Belgium
[3] Tech Univ Crete, Sch Elect & Comp Engn, Khania, Greece
[4] Univ Thessaly, Dept Biol, Fac Med, Larisa, Greece
[5] Ctr Res & Technol Hellas CERTH, Inst Res & Technol Thessaly, Larisa, Greece
[6] Univ Thessaly, Dept Orthoped, Fac Med, Larisa, Greece
关键词
Osteoarthritis; Cartilage; Chondrocytes; Proteomics; Mass spectrometry; Pathway analysis; PLS3; GSTP1; EPITHELIAL-MESENCHYMAL TRANSITION; PROTEIN EXPRESSION; ANDROGEN RECEPTOR; STATISTICAL-MODEL; GROWTH-PLATE; DIFFERENTIATION; CARTILAGE; BONE; PATHWAY; MS/MS;
D O I
10.1186/s12014-015-9085-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Background: Osteoarthritis (OA) is a multi-factorial disease leading progressively to loss of articular cartilage and subsequently to loss of joint function. While hypertrophy of chondrocytes is a physiological process implicated in the longitudinal growth of long bones, hypertrophy-like alterations in chondrocytes play a major role in OA. We performed a quantitative proteomic analysis in osteoarthritic and normal chondrocytes followed by functional analyses to investigate proteome changes and molecular pathways involved in OA pathogenesis. Methods: Chondrocytes were isolated from articular cartilage of ten patients with primary OA undergoing knee replacement surgery and six normal donors undergoing fracture repair surgery without history of joint disease and no OA clinical manifestations. We analyzed the proteome of chondrocytes using high resolution mass spectrometry and quantified it by label-free quantification and western blot analysis. We also used WebGestalt, a web-based enrichment tool for the functional annotation and pathway analysis of the differentially synthesized proteins, using the Wikipathways database. ClueGO, a Cytoscape plug-in, is also used to compare groups of proteins and to visualize the functionally organized Gene Ontology (GO) terms and pathways in the form of dynamical network structures. Results: The proteomic analysis led to the identification of a total of similar to 2400 proteins. 269 of them showed differential synthesis levels between the two groups. Using functional annotation, we found that proteins belonging to pathways associated with regulation of the actin cytoskeleton, EGF/EGFR, TGF-beta, MAPK signaling, integrin-mediated cell adhesion, and lipid metabolism were significantly enriched in the OA samples (p <= 10(-5)). We also observed that the proteins GSTP1, PLS3, MYOF, HSD17B12, PRDX2, APCS, PLA2G2A SERPINH1/HSP47 and MVP, show distinct synthesis levels, characteristic for OA or control chondrocytes. Conclusion: In this study we compared the quantitative changes in proteins synthesized in osteoarthritic compared to normal chondrocytes. We identified several pathways and proteins to be associated with OA chondrocytes. This study provides evidence for further testing on the molecular mechanism of the disease and also propose proteins as candidate markers of OA chondrocyte phenotype.
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页数:16
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