DNA demethylation is directly related to tumour progression: evidence in normal, pre-malignant and malignant cells from uterine cervix samples

A computer-assisted assay based on the quantitative analysis of DNA methylation in individual interphase nuclei by indirect immunolabelling with anti-5-methylcytosine antibodies was recently developed in our laboratory. In situ analyses were performed on individual nuclei from normal and experimenta...

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Bibliographic Details
Published in:	Oncology reports Vol. 10; no. 3; p. 545
Main Authors:	de Capoa, Adriana, Musolino, Angelina, Della Rosa, Simonetta, Caiafa, Paola, Mariani, Luciano, Del Nonno, Franca, Vocaturo, Amina, Donnorso, Raffaele Perrone, Niveleau, Alain, Grappelli, Claudio
Format:	Journal Article
Language:	English
Published:	Greece 01-05-2003
Subjects:	5-Methylcytosine - analysis 5-Methylcytosine - immunology Adult Antibodies, Monoclonal - analysis Carcinoma in Situ - genetics Carcinoma in Situ - metabolism Carcinoma in Situ - pathology Carcinoma, Squamous Cell - genetics Carcinoma, Squamous Cell - metabolism Carcinoma, Squamous Cell - pathology Cell Line Cervix Uteri - metabolism Cervix Uteri - pathology Disease Progression DNA Methylation DNA, Neoplasm - metabolism Epithelial Cells - metabolism Epithelial Cells - pathology Female Fluorescent Antibody Technique, Indirect Humans Image Processing, Computer-Assisted - methods In Situ Hybridization, Fluorescence Middle Aged Precancerous Conditions - genetics Precancerous Conditions - metabolism Precancerous Conditions - pathology Uterine Neoplasms - genetics Uterine Neoplasms - metabolism Uterine Neoplasms - pathology
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Summary:	A computer-assisted assay based on the quantitative analysis of DNA methylation in individual interphase nuclei by indirect immunolabelling with anti-5-methylcytosine antibodies was recently developed in our laboratory. In situ analyses were performed on individual nuclei from normal and experimentally hypo- or hypermethylated cultured cells as well as on human peripheral blood B-lymphocytes from normal and chronic lymphoid leukemia (CLL) samples. We present the results obtained on cells from patients affected by different degrees of preneoplastic or neoplastic changes of the uterine cervix as compared to normal controls. The analysis of DNA methylation in individual cells from cytofuge samples was performed as follows: within each nucleus the eu- and heterochromatin methylation levels were quantified in the grey scale range by dedicated software in terms of numbers, areas and optical densities (ODs) of the immunolabeled dense heterochromatic regions ("spots"), and of the optical density of nuclear background, i.e., of nuclear euchromatin. Analogously, in randomly chosen microscope fields of tissue sections from paraffin-embedded samples, progressive tissue demethylation was observed in dysplastic and cancer cells as compared to normal ones. Both methods showed significant and progressive DNA hypomethylation in dysplastic and cancer cells as compared to control specimens.
ISSN:	1021-335X