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| Status |
Public on Feb 11, 2010 |
| Title |
C-peptide and/or transforming growth factor beta 1 effect on human proximal tubular cell line |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Microarray analysis reveals up-regulation of retinoic acid and hepatocyte growth factor related signaling pathways by pro-insulin C-peptide in kidney proximal tubular cells: Antagonism of the pro-fibrotic effects of TGF-b1
Novel signaling roles for C-peptide have recently been discovered with evidence that it can ameliorate complications of type 1 diabetes. Here we sought to identify new pathways regulated by C-peptide of relevance to the pathophysiology of diabetic nephropathy. Microarray analysis was performed to identify genes regulated by either C-peptide and/or transforming growth factor beta 1 (TGF-β1) in a human proximal tubular cell line, HK-2. Expression of retinoic acid receptor β (RARβ), hepatcoyte growth factor (HGF), cellular retinoic acid binding protein II (CRABPII), vimentin, E-cadherin, Snail and β-catenin was assessed by immunoblotting. The cellular localisation of vimentin and β-catenin was determined by immunocytochemistry. Changes in cell morphology were assessed by phase contrast microscopy. Gene expression profiling demonstrated differential expression of 953 and 1,458 genes after C-peptide exposure for 18h or 48h respectively. From these, members of the anti-fibrotic retinoic acid (RA) and HGF signaling pathways were selected. Immunoblotting demonstrated that C-peptide increased RARβ, CRABPII and HGF. We confirmed a role for RA in reversal of TGF-β1-induced changes associated with epithelial-mesenchymal transition (EMT), including expression changes in Snail, E-cadherin, vimetin and redistribution of β-catenin. Importantly, these TGF-β1-induced changes were inhibited by C-peptide. Further, effects of TGF-β1 on Snail and E-cadherin expression were blocked by HGF and inhibitory effects of C-peptide were removed by blockade of HGF activity. This study identifies a novel role for HGF as an effector of C-peptide, possibly via an RA signaling pathway, highlighting C-peptide as a potential therapy for diabetic nephropathy.
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| Overall design |
For microarray analyses, all treatments were performed in triplicate to yield 18 flasks, and RNA from each flask hybridized to a separate chip to give an n of 3 for each of 6 treatments. Flasks were subjected to identical media changes and cells cultured for identical periods in media without supplements. In all experiments, cells were serum-starved overnight before agonist addition. Treatments were initiated such that 18h and 48h incubation periods ended coincidentally and all RNA was prepared at this point.
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| Contributor(s) |
Hills CE, Willars GB, Brunskill NJ |
| Citation(s) |
20197308 |
| Submission date |
Feb 09, 2010 |
| Last update date |
Feb 18, 2019 |
| Contact name |
Nigel John Brunskill |
| E-mail(s) |
njb18@le.ac.uk
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| Phone |
+44 116 2588043
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| Organization name |
University of Leicester
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| Department |
Infection Immunity and Inflammation
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| Street address |
University Road
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| City |
Leicester |
| State/province |
Leics |
| ZIP/Postal code |
LE1 9HN |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL6884 |
Illumina HumanWG-6 v3.0 expression beadchip |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA124211 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE20247_RAW.tar |
6.3 Mb |
(http)(custom) |
TAR |
| GSE20247_non-normalized_Sample_Probe_Profile_BSonly.txt.gz |
9.2 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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