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 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 microg/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 beta-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).
Singhto
Nilubon
N
Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Sintiprungrat
Kitisak
K
Sinchaikul
Supachok
S
Chen
Shui-Tein
ST
Thongboonkerd
Visith
V
D015153:Blotting, Western; D002129:Calcium Oxalate / Q000633:toxicity*; D015180:Electrophoresis, Gel, Two-Dimensional; D005453:Fluorescence; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D008954:Models, Biological; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry; D020298:U937 Cells
D002129:Calcium Oxalate