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

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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BMC Medical Genomics, BioMed Central, 2011, 4 (1), pp.39. 〈10.1186/1755-8794-4-39〉
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Soumis le : vendredi 27 janvier 2012 - 14:00:53
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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