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      Differential Proteome Analysis of Bone Marrow Mesenchymal Stem Cells from Adolescent Idiopathic Scoliosis Patients

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          Abstract

          Adolescent idiopathic scoliosis (AIS) is a complex three-dimensional deformity of the spine. The cause and pathogenesis of scoliosis and the accompanying generalized osteopenia remain unclear despite decades of extensive research. In this study, we utilized two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry (MS) to analyze the differential proteome of bone marrow mesenchymal stem cells (BM-MSCs) from AIS patients. In total, 41 significantly altered protein spots were detected, of which 34 spots were identified by MALDI-TOF/TOF analysis and found to represent 25 distinct gene products. Among these proteins, five related to bone growth and development, including pyruvate kinase M2, annexin A2, heat shock 27 kDa protein, γ-actin, and β-actin, were found to be dysregulated and therefore selected for further validation by Western blot analysis. At the protein level, our results supported the previous hypothesis that decreased osteogenic differentiation ability of MSCs is one of the mechanisms leading to osteopenia in AIS. In summary, we analyzed the differential BM-MSCs proteome of AIS patients for the first time, which may help to elucidate the underlying molecular mechanisms of bone loss in AIS and also increase understanding of the etiology and pathogenesis of AIS.

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          Most cited references75

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          Pyruvate kinase type M2 and its role in tumor growth and spreading.

          Proliferating cells and tumor cells in particular express the pyruvate kinase isoenzyme type M2 (M2-PK). Within the tumor metabolome M2-PK regulates the proportions of glucose carbons that are channelled to synthetic processes (inactive dimeric form) or used for glycolytic energy production (highly active tetrameric form, a component of the glycolytic enzyme complex). In tumor cells, the dimeric form of M2-PK (Tumor M2-PK) is always predominant. The dimerization is caused by direct interaction of M2-PK with certain oncoproteins. The switch between the tetrameric and dimeric form of M2-PK allows tumor cells to survive in environments with varying oxygen und nutrient supply.
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            The prediction of curve progression in untreated idiopathic scoliosis during growth.

            We reviewed the cases of 727 patients with idiopathic scoliosis in whom the initial curve measured from 5 to 29 degrees. The patients were followed either to the end of skeletal growth or until the curve progressed. One hundred and sixty-nine patients (23.2 per cent) showed progression of the curve. The incidence of curve progression was found to be related to the pattern and magnitude of the curve, the patient's age at presentation, the Risser sign, and the patient's menarchal status. We found no correlation between progression of the curve and the patient's sex, Harrington factor, rotational prominence, family history, or radiographic measurements. A progression factor was calculated using the three strongest correlations available at initial examination: the magnitude of the curve, the Risser sign, and the patient's chronological age. A graph and nomogram are presented that can serve as a guide for advising patients' families and for planning continuing care.
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              Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis.

              Resistance to stress-induced apoptosis was examined in cells in which the expression of hsp70 was either constitutively elevated or inducible by a tetracycline-regulated transactivator. Heat-induced apoptosis was blocked in hsp70-expressing cells, and this was associated with reduced cleavage of the common death substrate protein poly(ADP-ribose) polymerase (PARP). Heat-induced cell death was correlated with the activation of the stress-activated protein kinase SAPK/JNK (c-Jun N-terminal kinase). Activation of SAPK/JNK was strongly inhibited in cells in which hsp70 was induced to a high level, indicating that hsp70 is able to block apoptosis by inhibiting signaling events upstream of SAPK/JNK activation. In contrast, SAPK/JNK activation was not inhibited by heat shock in cells with constitutively elevated levels of hsp70. Cells that constitutively overexpress hsp70 resist apoptosis induced by ceramide, a lipid signaling molecule that is generated by apoptosis-inducing treatments and is linked to SAPK/JNK activation. Similar to heat stress, resistance to ceramide-induced apoptosis occurs in spite of strong SAPK/JNK activation. Therefore, hsp70 is also able to inhibit apoptosis at some point downstream of SAPK/JNK activation. Since PARP cleavage is prevented in both cell lines, these results suggest that hsp70 is able to prevent the effector steps of apoptotic cell death. Processing of the CED-3-related protease caspase-3 (CPP32/Yama/apopain) is inhibited in hsp70-expressing cells; however, the activity of the mature enzyme is not affected by hsp70 in vitro. Caspase processing may represent a critical heat-sensitive target leading to cell death that is inhibited by the chaperoning function of hsp70. The inhibition of SAPK/JNK signaling and apoptotic protease effector steps by hsp70 likely contributes to the resistance to stress-induced apoptosis seen in transiently induced thermotolerance.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2011
                22 April 2011
                : 6
                : 4
                : e18834
                Affiliations
                [1 ]Department of Orthopedics, Peking Union Medical College Hospital, Beijing, People's Republic of China
                [2 ]Center of Excellence in Tissue Engineering, Institute of Basic Medical Sciences and School of Basic Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People's Republic of China
                [3 ]State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, People's Republic of China
                Instituto Nacional de Câncer, Brazil
                Author notes

                Conceived and designed the experiments: GQ RCZ QZ ZW YJ. Performed the experiments: QZ JL TL YY. Analyzed the data: QZ JL WS YY. Contributed reagents/materials/analysis tools: JL JZ YY. Wrote the paper: QZ JL WS.

                Article
                PONE-D-11-03448
                10.1371/journal.pone.0018834
                3081308
                21526124
                ad5f9dc6-c910-4315-9ac9-675a477749bf
                Zhuang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 10 February 2011
                : 11 March 2011
                Page count
                Pages: 14
                Categories
                Research Article
                Biology
                Biochemistry
                Proteins
                Cytoskeletal Proteins
                Proteome
                Developmental Biology
                Stem Cells
                Mesenchymal Stem Cells
                Cell Differentiation
                Molecular Cell Biology
                Cellular Types
                Stem Cells
                Mesenchymal Stem Cells
                Proteomics
                Medicine
                Anatomy and Physiology
                Musculoskeletal System
                Bone
                Surgery
                Orthopedic Surgery
                Women's Health
                Osteopenia and Osteoporosis

                Uncategorized
                Uncategorized

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