Por favor, use este identificador para citar o enlazar este Item:http://hdl.handle.net/20.500.12105/10797
Título
Quantification of unmethylated Alu (QUAlu): a tool to assess global hypomethylation in routine clinical samples.
Autor(es)
Buj, Raquel | Mallona, Izaskun | Díez-Villanueva, Anna | Barrera, Víctor | Mauricio, Dídac | Puig-Domingo, Manel | Reverter, Jordi L | Matias-Guiu, Xavier | Azuara, Daniel | Alonso, Sergio | Rosell, Rafael | Capellà, Gabriel | Perucho, Manuel | Robledo Batanero, Mercedes CNIO | Peinado, Miguel A | Jordà, Mireia | Reverter, Jordi L. | Ramírez, Jose L | Peinado, Miguel A.
Fecha de publicación
2016-03-01
Cita
Oncotarget .2016 ;7(9):10536-46.
Idioma
Inglés
Tipo de documento
journal article
Resumen
Hypomethylation of DNA is a hallmark of cancer and its analysis as tumor biomarker has been proposed, but its determination in clinical settings is hampered by lack of standardized methodologies. Here, we present QUAlu (Quantification of Unmethylated Alu), a new technique to estimate the Percentage of UnMethylated Alu (PUMA) as a surrogate for global hypomethylation. QUAlu consists in the measurement by qPCR of Alu repeats after digestion of genomic DNA with isoschizomers with differential sensitivity to DNA methylation. QUAlu performance has been evaluated for reproducibility, trueness and specificity, and validated by deep sequencing. As a proof of use, QUAlu has been applied to a broad variety of pathological examination specimens covering five cancer types. Major findings of the preliminary application of QUAlu to clinical samples include: (1) all normal tissues displayed similar PUMA; (2) tumors showed variable PUMA with the highest levels in lung and colon and the lowest in thyroid cancer; (3) stools from colon cancer patients presented higher PUMA than those from control individuals; (4) lung squamous cell carcinomas showed higher PUMA than lung adenocarcinomas, and an increasing hypomethylation trend associated with smoking habits. In conclusion, QUAlu is a simple and robust method to determine Alu hypomethylation in human biospecimens and may be easily implemented in research and clinical settings.
MESH
Adenocarcinoma | Adenocarcinoma of Lung | Alu Elements | Biomarkers, Tumor | Carcinoma, Squamous Cell | Cell Line, Tumor | Colonic Neoplasms | CpG Islands | DNA | DNA Methylation | HCT116 Cells | Humans | Lung Neoplasms | Molecular Diagnostic Techniques | Polymerase Chain Reaction | Thyroid Neoplasms
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