Down-Regulation of Connexin 32 Gene Expression through DNA Methylation in a Human Renal Cell Carcinoma Cell

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.

Journal

Journal ID (publisher-id): AJN

Journal ID (nlm-ta): Am J Nephrol

Journal ID (doi): 10.1159/issn.0250-8095

Title: American Journal of Nephrology

Publisher: S. Karger AG

ISSN (Print): 0250-8095

ISSN (Electronic): 1421-9670

Publication date Subscription-year: 2003

Publication date (Print): June 2003

Publication date (Electronic): 16 May 2003

Volume: 23

Issue: 3

Pages: 172-177

Affiliations

^aDepartment of Veterinary Pharmacology, College of Bioresource Sciences, Nihon University, Kanagawa; ^bDepartment of Food Science Research for Health, National Institute of Health and Nutrition, Tokyo, and ^cKwansei Gaskuin University School of Science and Technology, Hyogo, Japan

Article

Publisher ID: 70653 Other ID: Am J Nephrol 2003;23:172–177

DOI: 10.1159/000070653

PubMed ID: 12690227

Copyright statement: © 2003 S. Karger AG, Basel

License:

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Page count

Figures: 2, References: 19, Pages: 6

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Categories

Subject: Original Article: Basic Sciences