%0 Journal Article %T Detecting differential allelic expression using high-resolution melting curve analysis: application to the breast cancer susceptibility gene CHEK2. %+ Section Génétique - Groupe Prédispositions génétiques au cancer %+ Oncogénèse et progression tumorale %+ Unité Mixte de Génétique Constitutionnelle des Cancers Fréquents %+ Department of Pathology %+ Department of Oncological Sciences %A Nguyen-Dumont, Tú %A Jordheim, Lars %A Michelon, Jocelyne %A Forey, Nathalie %A Mckay-Chopin, Sandrine %A Sinilnikova, Olga %A Le Calvez-Kelm, Florence %A Southey, Melissa %A Tavtigian, Sean %A Lesueur, Fabienne %Z kConFab is supported by grants from the National Breast Cancer Foundation, the National Health and Medical Research Council (NHMRC) and by the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia. We gratefully acknowledge Arnaud Dumont for his help on developing the R script. TN-D was the recipient of a fellowship from Fondation de France and a Special Trainee Award from the International Agency for Research on Cancer. LPJ was the recipient of a fellowship from Ligue Contre le Cancer - Comité du Rhône. MCS is a National Health and Medical Research Council Senior Research Fellow and Victorian Breast cancer Research Consortium (VBCRC) Group Leader. This work was also supported by the Canadian Institutes of Health Research team grant CRN-87521-IC089832 and by the National Cancer Institute, National Institutes of Health (NIH) grant R01 CA121245. %< avec comité de lecture %@ 1755-8794 %J BMC Medical Genomics %I BioMed Central %V 4 %N 1 %P 39 %8 2011 %D 2011 %R 10.1186/1755-8794-4-39 %M 21569354 %Z Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN]Journal articles %X 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. %G English %Z for Kathleen Cuningham Foundation Consortium for Research into Familial Aspects of Breast Cancer (kConFab) %2 https://inserm.hal.science/inserm-00663661/document %2 https://inserm.hal.science/inserm-00663661/file/1755-8794-4-39.pdf %L inserm-00663661 %U https://inserm.hal.science/inserm-00663661 %~ INSERM %~ HCL %~ UNIV-LYON1 %~ UDL %~ UNIV-LYON