Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2.

Tú Nguyen-Dumont 1 Lars Jordheim 2 Jocelyne Michelon 1 Nathalie Forey 1 Sandrine Mckay-Chopin 1 Olga Sinilnikova 3 Florence Le Calvez-Kelm 1 Melissa Southey 4 Sean Tavtigian 5 Fabienne Lesueur 1, *
* Auteur correspondant
1 Section Génétique - Groupe Prédispositions génétiques au cancer
2 Oncogénèse et progression tumorale
3 Unité Mixte de Génétique Constitutionnelle des Cancers Fréquents
4 Department of Pathology
5 Department of Oncological Sciences
Abstract : BACKGROUND: The gene CHEK2 encodes a checkpoint kinase playing a key role in the DNA damage pathway. Though CHEK2 has been identified as an intermediate breast cancer susceptibility gene, only a small proportion of high-risk families have been explained by genetic variants located in its coding region. Alteration in gene expression regulation provides a potential mechanism for generating disease susceptibility. The detection of differential allelic expression (DAE) represents a sensitive assay to direct the search for a functional sequence variant within the transcriptional regulatory elements of a candidate gene. We aimed to assess whether CHEK2 was subject to DAE in lymphoblastoid cell lines (LCLs) from high-risk breast cancer patients for whom no mutation in BRCA1 or BRCA2 had been identified. METHODS: We implemented an assay based on high-resolution melting (HRM) curve analysis and developed an analysis tool for DAE assessment. RESULTS: We observed allelic expression imbalance in 4 of the 41 LCLs examined. All four were carriers of the truncating mutation 1100delC. We confirmed previous findings that this mutation induces non-sense mediated mRNA decay. In our series, we ruled out the possibility of a functional sequence variant located in the promoter region or in a regulatory element of CHEK2 that would lead to DAE in the transcriptional regulatory milieu of freely proliferating LCLs. CONCLUSIONS: Our results support that HRM is a sensitive and accurate method for DAE assessment. This approach would be of great interest for high-throughput mutation screening projects aiming to identify genes carrying functional regulatory polymorphisms.
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Article dans une revue
BMC Medical Genomics, BioMed Central, 2011, 4 (1), pp.39. 〈10.1186/1755-8794-4-39〉
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Dernière modification le : jeudi 8 février 2018 - 11:08:23
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Tú Nguyen-Dumont, Lars Jordheim, Jocelyne Michelon, Nathalie Forey, Sandrine Mckay-Chopin, et al.. Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2.. BMC Medical Genomics, BioMed Central, 2011, 4 (1), pp.39. 〈10.1186/1755-8794-4-39〉. 〈inserm-00663661〉

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