Abstract
BackgroundCalcium oxalate monohydrate (COM), the major crystalline composition of most kidney stones, induces inflammatory infiltration and injures in renal tubular cells. However, the mechanism of COM-induced toxic effects in renal tubular cells remain ambiguous. The present study aimed to investigate the potential changes in proteomic landscape of proximal renal tubular cells in response to the stimulation of COM crystals.MethodsClinical kidney stone samples were collected and characterized by a stone component analyzer. Three COM-enriched samples were applied to treat human proximal tubular epithelial cells HK-2. The proteomic landscape of COM-crystal treated HK-2 cells was screened by TMT-labeled quantitative proteomics analysis. The differentially expressed proteins (DEPs) were identified by pair-wise analysis. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEPs were performed. Protein interaction networks were identified by STRING database.ResultsThe data of TMT-labeled quantitative proteomic analysis showed that a total of 1141 proteins were differentially expressed in HK-2 cells, of which 699 were up-regulated and 442 were down-regulated. Functional characterization by KEGG, along with GO enrichments, suggests that the DEPs are mainly involved in cellular components and cellular processes, including regulation of actin cytoskeleton, tight junction and focal adhesion. 3 high-degree hub nodes, CFL1, ACTN and MYH9 were identified by STRING analysis.ConclusionThese results suggested that calcium oxalate crystal has a significant effect on protein expression profile in human proximal renal tubular epithelial cells.
Highlights
Calcium oxalate monohydrate (COM), the major crystalline composition of most kidney stones, induces inflammatory infiltration and injures in renal tubular cells
We aim to use TMT (Tandem Mass Tag)–based quantitative proteomics analysis to investigate the effects of calcium oxalate crystal on the differential protein expression profiles of human renal tubular epithelial cell HK-2, screen differentially expressed protein molecules and initially explore their functional roles in calcium oxalate stone formation and its resulting renal damage
Preparation of kidney stone treated HK-2 cells and quality control of TMT mass spectrometry samples The human proximal tubular epithelial cell HK-2 cells were suspended into single cells, seeded onto 6-well plates overnight
Summary
Calcium oxalate monohydrate (COM), the major crystalline composition of most kidney stones, induces inflammatory infiltration and injures in renal tubular cells. The mechanism of COM-induced toxic effects in renal tubular cells remain ambiguous. Kidney stone is a common disease with prevalence and incidence increasing across the world that seriously affects human health. It can cause urinary retention, renal pelvis and ureteral hydrops, ureteral dilatation, renal function damage, and infection. The prevalence of kidney stones exhibited a significant increasing trend in the past decades in the mainland of China [1, 2]. Kidney stone seriously affects the health and quality of life of patients, and has become a public health problem across the world
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