Résumé
L’hybridation génomique comparée (HGC) est une technique developpée récemment qui permet, en une seule analyse, la détection d’aberrations génétiques pour la totalité du génome de la tumeur sans nécessiter de culture cellulaire; elle peut s’appliquer à du matériel conservé pour archive. Elle procure une vision globale du gain ou de la perte d’ADN dans une tumeur en comparaison à de l’ADN contrôle. Les pertes peuvent correspondre à des gènes suppresseurs de tumeurs connus ou non et les gains à l’amplification d’oncogènes, les uns et les autres impliqués dans la pathogénèse du cancer colorectal. Au cours de cette procédure, l’ADN test obtenu à partir d’échantillons de tumeurs est marqué avec un fluorochrome vert et mélangé avec de l’ADN contrôle marqué en rouge et obtenu à partir de cellules avec un contenu chromosomique diploïde normal. Cette mixture est hybridée à des préparations métaphasiques normales. L’hybridation des séquences d’ADN test ou contrôle, qui rentrent en compétition lors de l’appariement, est reconnue grâce aux différents fluorochromes. Le ratio de fluorescence verte par rapport à rouge pour chaque région chromosomique, obtenu par analyse d’image digitale, est la mesure de la sous-ou sur-représentation de matériel génétique pour cette région de la tumeur étudiée.
Comparée à d’autres techniques, l’HGC parvient à une information plus spécifique que, par exemple, l’analyse de l’ADN en cytométrie de flux, et à une meilleure vision globale des modifications génétiques au sein d’une tumeur qu’avec l’hybridationin situ ou l’analyse de perte d’hétérozygotie. Les applications de l’hybridation génomique comparative sont multiples, par exemple pour le criblage d’aberrations chromosomiques dans une large série de tumeurs destiné à identifier des loci chromosomiques qui renferment possiblement de nouveaux gènes suppresseurs de tumeurs ou de nouveaux oncogènes, pour l’analyse génétique des modifications au cours des stades successifs de la carcinogénèse, ou pour élucider les bases génétiques des caractéristiques cliniques et phénotypiques de certaines tumeurs.
Summary
Comparative Genomic Hybridization (CGH) is a recently developed technique that allows to detect in one analysis genetic aberrations in the complete tumour genome without the need of cell culturing, and it can be applied to archival material. It provides an overview of chromosomal losses and gains of DNA in a tumour as compared to control DNA. Losses can point to known and putative tumour suppressor genes and gains to amplification of oncogenes, involved in the pathogenesis of colorectal cancer. In this procedure, test DNA obtained from tumour specimens is labelled with a green fluorochrome and mixed with red labelled control DNA obtained from cells with a normal diploid chromosome complement. This mixture is hybridized to normal metaphase preparations. Hybridized test and control DNA sequences, which compete in the annealing process, are recognized via the different fluorochromes. The ratio of green to red fluorescence for each chromosomal region, analysed by digital image processing, is a measure of the under- or over-representation of genetic material for that region in the tumour studied.
Compared to other techniques, CGH yields more specific information than e.g. DNA cytometry, and a better overview of genetic changes in a tumour than is possible with in situ hybridization or loss of heterozygosity analysis. The applications of comparative genomic hybridization are multiple, e.g. to screen for chromosomal aberrations in large series of tumours aiming to identify chromosomal loci that possibly harbour new tumour suppressor genes or oncogenes, to analyse genetic changes in subsequent stages of carcinogenesis, or to elucidate the genetic basis of clinical and phenotypical tumour characteristics.
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Meijer, G.A., Aja Hermsen, M., Meuwissen, S.G. et al. L’hybridation génomique comparative ouvre de nouvelles perspectives dans l’exploration des modifications qui accompagnent les lésions (pré-)malignes du tube digestif. Acta Endosc 27, 89–99 (1997). https://doi.org/10.1007/BF02968930
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DOI: https://doi.org/10.1007/BF02968930