Proteome Changes in Human Monocytes upon Interaction with Calcium Oxalate Monohydrate Crystals

Monocytic infiltration in renal interstitium is commonly found surrounding the site of calcium oxalate (CaOx) crystal deposition in the kidney. Monocytes are supposed to eliminate the deposited crystals. However, effects of CaOx crystals on the infiltrating monocytes remain unknown. Therefore, this...

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Bibliographic Details
Published in:Journal of proteome research 2010-08, Vol.9 (8), p.3980-3988
Main Authors: Singhto, Nilubon, Sintiprungrat, Kitisak, Sinchaikul, Supachok, Chen, Shui-Tein, Thongboonkerd, Visith
Format: Article
Language:English
Subjects:
Blotting, Western
Calcium Oxalate - toxicity
Electrophoresis, Gel, Two-Dimensional
Fluorescence
Gene Expression Regulation - drug effects
Humans
Models, Biological
Monocytes - drug effects
Monocytes - metabolism
Oxidative Stress - drug effects
Proteomics - methods
Tandem Mass Spectrometry
U937 Cells
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Summary:Monocytic infiltration in renal interstitium is commonly found surrounding the site of calcium oxalate (CaOx) crystal deposition in the kidney. Monocytes are supposed to eliminate the deposited crystals. However, effects of CaOx crystals on the infiltrating monocytes remain unknown. Therefore, this study investigated the altered cellular proteome of human monocytes in response to interaction with CaOx monohydrate (COM) crystals. After 24-h culture with or without 100 μg/mL COM crystals, U937 cells were harvested and subjected to 2-DE analysis with Deep Purple fluorescence staining (n = 5 gels/group; each was derived from independent culture). Spot matching, quantitative intensity analysis, and statistics revealed 22 differentially expressed proteins (9 up-regulated and 13 down-regulated proteins), which were successfully identified by Q-TOF MS and MS/MS analyses, including those involved in cell cycle, cellular structure, carbohydrate metabolism, lipid metabolism, mRNA processing, and protein synthesis, stabilization, and degradation. Randomly selected changes [up-regulated ALG-2 interacting protein 1 (Alix), elongation factor-2 (EF-2), and down-regulated β-actin] were confirmed by Western blot analysis. Our data may help to understand how monocytes interact with COM crystals. These processes are proposed to cause subsequent inflammatory response in kidney stone disease through oxidative stress pathway(s).
ISSN:1535-3893
1535-3907
DOI:10.1021/pr100174a