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Am J Nephrol. 2003 May-Jun;23(3):172-7. Epub 2003 Apr 10.

Down-regulation of connexin 32 gene expression through DNA methylation in a human renal cell carcinoma cell.

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  • 1Department of Veterinary Pharmacology, College of Bioresource Sciences, Nihon University, Kanagawa, Japan.

Abstract

BACKGROUND:

We have recently reported that connexin (Cx) 32 is down-regulated in a human renal cell carcinoma (RCC) cell (Caki-2 cell).

HYPOTHESIS:

We postulated that the down-regulation of Cx32 gene in the RCC cell is due to hypermethylation of its promoter region.

METHODS:

We estimated methylation status in the promoter region of Cx32 gene in the RCC cell by DNA digestion with methylation-sensitive restriction enzyme and PCR, and methylation-specific PCR (MSP). We also checked the recovery of Cx32 gene expression in the RCC cell treated with a DNA methyltranferase inhibitor, 5-Aza-2'-deoxycytidine (5-Aza-CdR).

RESULTS:

Treatment with 5-Aza-CdR resulted in induction of Cx32 expression in the RCC cell. Hypermethylation of the Cx32 promoter region in the RCC cell was confirmed by DNA digestion with methylation-sensitive restriction enzyme and PCR, and MSP.

CONCLUSION:

We suggest that hypermethylation in the promoter region is a mechanism for the Cx32 gene repression in the RCC cell.

Copyright 2003 S. Karger AG, Basel

PMID:
12690227
[PubMed - indexed for MEDLINE]
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