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Analyses of the genomic methylation status of the human cyclin A1 promoter by a novel real‐time PCR‐based methodology
Author(s) -
Müller-Tidow Carsten,
Bornemann Christina,
Diederichs Sven,
Westermann Annette,
Klümpen Silvia,
Zuo Peijun,
Wang Wenbing,
Berdel Wolfgang E,
Serve Hubert
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02128-7
Subject(s) - microbiology and biotechnology , dna methylation , biology , methylation , cpg site , bisulfite sequencing , illumina methylation assay , differentially methylated regions , cyclin a2 , genomic dna , cyclin d , gene expression , gene , cyclin , genetics , cell cycle
The role of CpG methylation in the regulation of tissue‐specific gene expression is highly controversial. Cyclin A1 is a tissue‐specifically expressed gene that is strongly methylated in non‐expressing tumor cell lines. We have established a novel real‐time PCR method to quantitate genomic CpG methylation of the cyclin A1 promoter. Genomic DNA samples from different human organs were treated with bisulfite and amplified with methylation‐specific primers and with primers amplifying methylated as well as non‐methylated DNA. PCR product quantitation was obtained by using a fluorogenic probe labeled with FAM and TAMRA. These analyses demonstrated that the human cyclin A1 promoter was methylated in kidney, colon, spleen, testis, and small intestine, but not in brain, liver, pancreas, or heart. Expression of cyclin A1 was predominantly found in testis. Low level expression of cyclin A1 was present in spleen, prostate, leukocytes, colon, and thymus. Taken together, our data provide evidence that CpG methylation patterns of the human cyclin A1 promoter in human organs do not generally correlate with cyclin A1 gene expression in vivo.