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
BACKGROUND: We have recently reported that connexin (Cx) 32 is down-regulated in a humanrenal 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
Authors: H Hagiwara; H Sato; Y Ohde; Y Takano; T Seki; T Ariga; N Hokaiwado; M Asamoto; T Shirai; Y Nagashima; T Yano Journal: Br J Pharmacol Date: 2008-02-11 Impact factor: 8.739