Methylation status of the Ep-CAM promoter region in human breast cancer cell lines and breast cancer tissue.
We examined the methylation status of the Ep-CAM promoter region of human breast cancer cell lines and breast cancer tissue using MethyLight technology and bisulfite sequencing. We found the promoter of Ep-CAM-negative breast cancer cell lines Hs 578T to be methylated to a higher degree as compared to positive cell lines MCF-7. Demethylation of cell lines was performed using 5-aza-2'-deoxycytidine. Ep-CAM RNA and protein expression could be partially restored by treating cells with 5-Aza-2'-deoxycytidine. In most primary breast cancer tissue, methylation of the Ep-CAM gene could be detected at a low level and no correlation was found with Ep-CAM protein expression in tumour tissue. Taken together, these data suggest that methylation of the Ep-CAM promoter is not a crucial mechanism for regulation of Ep-CAM expression in breast cancer. Thus, most important regulatory mechanisms have to be supposed in vivo.
Spizzo
Gilbert
G
Division of Haematology and Oncology, Innsbruck Medical University, Innsbruck, Austria. gilbert.spizzo@uibk.ac.at
Gastl
Guenther
G
Obrist
Peter
P
Fong
Dominic
D
Haun
Margot
M
Grünewald
Kurt
K
Parson
Walther
W
Eichmann
Cordula
C
Millinger
Simone
S
Fiegl
Heidi
H
Margreiter
Raimund
R
Amberger
Albert
A
D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D001940:Breast / Q000378:metabolism; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D003106:Colon / Q000378:metabolism; D018899:CpG Islands; D019175:DNA Methylation; D000077209:Decitabine; D000071858:Epithelial Cell Adhesion Molecule; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D012333:RNA, Messenger; D000077209:Decitabine; D001374:Azacitidine