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

Am J Nephrol. 2003 May-Jun;23(3):172-7. doi: 10.1159/000070653. Epub 2003 Apr 10.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Azacitidine / pharmacology
  • Carcinoma, Renal Cell / genetics*
  • Cell Communication
  • Connexins / genetics*
  • DNA Methylation*
  • Down-Regulation
  • Gap Junction beta-1 Protein
  • Gene Expression*
  • Humans
  • Immunoblotting
  • Kidney Neoplasms / genetics*
  • Promoter Regions, Genetic
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tumor Cells, Cultured

Substances

  • Connexins
  • Azacitidine