摘要
AIM:To study the role of hypermethylation in the loss of retinoic acid receptor β2(RARβ2) in esophageal squamous cell carcinoma (ESCC).METHODS:The role of hypermethylation in RARβ2 gene silencing in 6 ESCC cell lines was determined by methylation-specific PCR (MSP),and its methylation status was compared with RARβ2 mRNA expression by RT-PCR.The MSP results were confirmed by bisulfite sequencing of RARβ2promoter regions.RESULTS:Methylation was detected in 4 of the 6 cell lines,and the expression of RARβ2 was markedly downregulated in 3 of the 4 methylated cell lines. The expression of RARβ2 was restored in one RARβ2-downregulated cell line with the partial demethylation of promoter region of RARβ2 after 5-aza-2′-deoxycytidine (5-aza-dc) treatment.CONCLUSION:The methylation of the 5′ region may play an important role in the downregulation of RARβ2 in some ESCC cell lines, suggesting that multiple mechanisms contribute to the loss of RARβ2expression in ESCC cell lines.This study may have clinical applications for treatment and prevention of ESCC.
AIM.To study the role of hypermethylation in the loss of retinoic acid receptor β_2(RARβ_2) in esophageal squamous cell carcinoma (ESCC). METHODS:The role of hypermethylation in RARβ_2 gene silencing in 6 ESCC cell lines was determined by methylation- specific PCR (MSP),and its methylation status was compared with RARβ_2 mRNA expression by RT-PCR.The MSP results were confirmed by bisulfite sequencing of RARβ_2 promoter regions. RESULTS:Methylation was detected in 4 of the 6 cell lines, and the expression of RARβ_2 was markedly downregulated in 3 of the 4 methylated cell lines.The expression of RARβ_2 was restored in one RARβ_2-downregulated cell line with the partial demethylation of promoter region of RARβ_2 after 5- aza-2'-deoxycytidine (5-aza-dc) treatment. CONCLUSION:The methylation of the 5' region may play an important role in the downregulation of RARβ_2 in some ESCC cell lines,suggesting that multiple mechanisms contribute to the loss of RARβ_2 expression in ESCC cell lines. This study may have clinical applications for treatment and prevention of ESCC.
基金
Supported by China Key Program on Basic Research,G1998051021
the Chinese Hi-tech R&D program,2001AA231041
National Science Foundation of China,30170519
