AIM: To verify the expression and methylation status o the MAGE-A1 and MAGE-A3 genes in colorectal cance tissues and cancer cell lines. METHODS: We evaluated promoter demethylation status of the MAGE-A1 and MAGE-A3 ge...AIM: To verify the expression and methylation status o the MAGE-A1 and MAGE-A3 genes in colorectal cance tissues and cancer cell lines. METHODS: We evaluated promoter demethylation status of the MAGE-A1 and MAGE-A3 genes by RT-PCR analysis and methylation-specific PCR (MS-PCR), as we as sequencing analysis, after sodium bisulfite modifica tion in 32 colorectal cancer cell lines and 87 cancer tis sues. RESULTS: Of the 32 cell lines, MAGE-A1 and MAGE-A3 expressions were observed in 59% and 66%, respective ly. Subsequent to sodium bisulfite modification and MS PCR analysis, the promoter hypomethylation of MAGE-A1 and MAGE-A3 was confirmed in both at 81% each Promoter hypomethylation of MAGE-A1 and MAGE-A3 in colorectal cancer tissues was observed in 43% and 77% respectively. Hypomethylation of MAGE-A1 and MAGE-A3 genes in corresponding normal tissues were observed in 2% and 6%, respectively. CONCLUSION: The promoter hypomethylation of MAGE genes up-regulates its expression in colorectal carcino mas as well as in gastric cancers and might play a signif icant role in the development and progression of human colorectal carcinomas.展开更多
To understand the DNA-methylation mediated gene silencing mechanisms, we analyzed in cell culture of the promoter function of the MAGE-A1 gene, which is frequently demethylated and over-expressed in human hepatocellul...To understand the DNA-methylation mediated gene silencing mechanisms, we analyzed in cell culture of the promoter function of the MAGE-A1 gene, which is frequently demethylated and over-expressed in human hepatocellular carcinoma. We have established the correlation of the DNA methylation of the promoter CpG island with expression status of this gene in a panel of the established liver cancer cell lines. The crucial CpG dinucleotide(s) within the minimal promoter subjected to the control mediated by DNA methylation with profound biological functions was also delineated.Furthermore, a novel sequence-specific DNA-protein interaction at the -30 CpG dinucleotide upstream of the gene was found having a vital part to play in the DNA methylation mediated transcription silencing of the MAGE-A1 gene. Our results would not only provide new insights into the DNA methylation mediated mechanisms over transcription of the MAGE-A1 gene, but also pave the way for further defining the cross-talk among DNA methylation, histone modification and chromatin remodeling in detail.展开更多
基金the Korea Research Foundation Grant, No.KRF-2003-03-E00199
文摘AIM: To verify the expression and methylation status o the MAGE-A1 and MAGE-A3 genes in colorectal cance tissues and cancer cell lines. METHODS: We evaluated promoter demethylation status of the MAGE-A1 and MAGE-A3 genes by RT-PCR analysis and methylation-specific PCR (MS-PCR), as we as sequencing analysis, after sodium bisulfite modifica tion in 32 colorectal cancer cell lines and 87 cancer tis sues. RESULTS: Of the 32 cell lines, MAGE-A1 and MAGE-A3 expressions were observed in 59% and 66%, respective ly. Subsequent to sodium bisulfite modification and MS PCR analysis, the promoter hypomethylation of MAGE-A1 and MAGE-A3 was confirmed in both at 81% each Promoter hypomethylation of MAGE-A1 and MAGE-A3 in colorectal cancer tissues was observed in 43% and 77% respectively. Hypomethylation of MAGE-A1 and MAGE-A3 genes in corresponding normal tissues were observed in 2% and 6%, respectively. CONCLUSION: The promoter hypomethylation of MAGE genes up-regulates its expression in colorectal carcino mas as well as in gastric cancers and might play a signif icant role in the development and progression of human colorectal carcinomas.
基金supported by the National High Technology Research and Development Program of China(863 Program)(2002AA2Z3352)the Major State Basic Research Development Program of China(973 Program)(G1998051004)the Science Foundation of Shanghai Municipal Government(02DJ14056)to Jing De ZHU.
文摘To understand the DNA-methylation mediated gene silencing mechanisms, we analyzed in cell culture of the promoter function of the MAGE-A1 gene, which is frequently demethylated and over-expressed in human hepatocellular carcinoma. We have established the correlation of the DNA methylation of the promoter CpG island with expression status of this gene in a panel of the established liver cancer cell lines. The crucial CpG dinucleotide(s) within the minimal promoter subjected to the control mediated by DNA methylation with profound biological functions was also delineated.Furthermore, a novel sequence-specific DNA-protein interaction at the -30 CpG dinucleotide upstream of the gene was found having a vital part to play in the DNA methylation mediated transcription silencing of the MAGE-A1 gene. Our results would not only provide new insights into the DNA methylation mediated mechanisms over transcription of the MAGE-A1 gene, but also pave the way for further defining the cross-talk among DNA methylation, histone modification and chromatin remodeling in detail.
