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Biochem Biophys Res Commun. 1997 Sep 8;238(1):179-84.

Identification of glucose-regulated genes in human mesangial cells by mRNA differential display.

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Division of Biomedical Sciences, Imperial College School of Medicine, Charing Cross Hospital, London, United Kingdom.


Diabetic nephropathy is characterised by an accumulation of extracellular matrix proteins in the glomerular mesangium. Hyperglycaemia is a major factor promoting this progressive expansion of the mesangial matrix. We have used the technique of mRNA differential display to investigate changes in gene expression in cultured human mesangial cells following long-term (21 days) exposure to either physiologic (4 mM) or pathologic (30 mM) D-glucose concentrations. Approximately 12,000 mRNA species were screened for evidence of altered expression and several hundred candidate cDNA fragments were obtained. Northern blot and RT-PCR analysis of ten randomly chosen candidate cDNA fragments revealed three exhibiting increased mRNA expression under elevated D-glucose levels. Nucleotide sequence analysis identified two of the cDNA fragments as encoding prolyl 4-hydroxylase alpha-subunit and thrombospondin-1. The third cDNA fragment represents a novel glucose-regulated gene, encoding a putative zinc finger protein. Upregulated expression of these genes in response to high levels of D-glucose may contribute significantly to the disease process. mRNA differential display is a useful tool to investigate the mechanism of diabetic nephropathy.

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