Hypermethylation of calcitonin gene regulatory sequences in human breast cancer as revealed by genomic sequencing.
DNA methylation has been studied intensively during the past years in order to elucidate its role in the regulation of gene expression, gene imprinting and cancer progression. Earlier studies have shown that a general genomic under-methylation is associated with chronic lymphocytic leukemia and metastatic prostate cancer. Site-specific methylation changes, as revealed by the use of methylation-sensitive restriction enzymes, have been reported to occur in the promotor region of the calcitonin gene in chronic myeloid leukemia as it progresses from the chronic phase to blast crisis, in non-Hodgkin's lymphoid neoplasms and in non-lymphocytic leukemia. We have now explored possible methylation changes associated with benign and malignant breast tumors. Two approaches were employed: (i) chemical determination of general genomic methylation status and (ii) base-specific analysis of the methylation changes in the promoter of the calcitonin gene with the aid of genomic sequencing. The results did not reveal any changes of total DNA 5-methylcytosine content in ductal carcinoma of breast in comparison with benign tumors. There was a small, yet significant, increase in 5-methylcytosine content in lobular carcinoma. Genomic sequencing of the promoter region of the calcitonin gene, however, revealed a striking hypermethylation at or around the transcription start site of the gene in ductal carcinomas. In benign tumors and lobular carcinomas, this region was either entirely unmethylated or only slightly methylated. The latter changes may reflect a regional hypermethylation of the short arm of chromosome 11, which harbors, in addition to the calcitonin gene, a number of putative or established tumor-suppressor genes. Our results demonstrate that genomic sequencing in its present form can be used for a reliable and precise DNA methylation analysis of primary human tumors.
Hakkarainen
M
M
A.I. Virtanen Institute, University of Kuopio, Finland.
Wahlfors
J
J
Myöhänen
S
S
Hiltunen
M O
MO
Eskelinen
M
M
Johansson
R
R
Jänne
J
J
D044503:5-Methylcytosine; D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics*; D002116:Calcitonin / Q000235:genetics*; D018270:Carcinoma, Ductal, Breast / Q000737:chemistry / Q000235:genetics; D018275:Carcinoma, Lobular / Q000737:chemistry / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003596:Cytosine / Q000031:analogs & derivatives / Q000032:analysis; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000737:chemistry*; D005260:Female; D018226:Fibroadenoma / Q000737:chemistry / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods
D004273:DNA, Neoplasm; D044503:5-Methylcytosine; D003596:Cytosine; D002116:Calcitonin