Résumé
Nous avons développé une technique sensible et reproductible d’hybridationin situ (HIS) en utilisant des sondes d’ARNc (ARN complémentaire) marquées à la digoxigénine pour localiser l’ARNm (ARM messager) de cytokératines (CK) sur des spécimens de muqueuse œsophagienne fixés ou inclus en paraffine, y compris des biopsies prélevées en endoscopie.
Comme le profil d’expression de CK est une caractéristique reconnue d’un type de cellule épithéliale donné ou d’un tissu, des changements spécifiques dans ce profil d’expression de CK ou dans l’expression d’ARNm-CK individuels peuvent être liés au développement précoce de pathologies épithéliales particulières telles que dysplasie et cancer. Etant donné que les modifications moléculaires précèdent souvent les changements morphologiques, la technique d’HIS décrite ici peut être extrapolée à la détection de modifications de l’expression de certains gènes et de la néo-expression d’autres marqueurs moléculaires, connus pour ou suspectés d’accompagner le processus cancéreux. En outre, ce protocole d’HIS bien établi peut être facilement réalisé et devenir un outil utile en histologie de routine, en recherche clinique ou pour le diagnostic clinique.
Summary
We developed a sensitive and reproducible in situ hybridisation (ISH) technique using digoxigenin-labelled cRNA probes for the localisation of cytokeratin (CK)-mRNA on fixed and parraffin-embedded human esophageal mucosa, including endoscopic biopsy specimens.
Since the CK pattern is accepted to be characteristic of a given epithelial cell or tissue, specific changes in the CK pattern or in the expression of individual CK-mRNAs may be related with the early development of particular epithelial pathologies, such as dysplasia and cancer. Since molecular changes often precede morphological changes, the ISH technique described may be extrapolated to detect changes in gene-expression or new expression of other molecular markers, known or suspected to accompany the onset of malignancy. Furthermore, the estabished ISH-protocol can be easily performed and may become a useful tool in routine histology, clinical research and clinical diagnosis.
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Viaene, A., Lerut, T. & Geboes, K. L’hybridationin situ: un outil polyvalent pour localiser l’expression de gènes sur sections tissulaires. Acta Endosc 27, 117–127 (1997). https://doi.org/10.1007/BF02968932
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DOI: https://doi.org/10.1007/BF02968932