Versatile Electroanalytical Bioplatforms for Simultaneous Determination of Cancer-Related DNA 5‑Methyl- and 5‑Hydroxymethyl-Cytosines at Global and Gene-Specific Levels in Human Serum and Tissues

• Published in: ACS sensors Vol. 4; no. 1; pp. 227 - 234
• Main Authors: Povedano, Eloy, Montiel, Víctor Ruiz-Valdepeñas, Valverde, Alejandro, Navarro-Villoslada, Fernando, Yáñez-Sedeño, Paloma, Pedrero, María, Montero-Calle, Ana, Barderas, Rodrigo, Peláez-García, Alberto, Mendiola, Marta, Hardisson, David, Feliú, Jaime, Camps, Jordi, Rodríguez-Tomàs, Elisabet, Joven, Jorge, Arenas, Meritxell, Campuzano, Susana, Pingarrón, José M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 25.01.2019
• Subjects: 5-Methylcytosine - analogs & derivatives; 5-Methylcytosine - blood; 5-Methylcytosine - immunology; Antibodies, Monoclonal - immunology; Armoracia - enzymology; Biomarkers, Tumor - blood; Biomarkers, Tumor - chemistry; Biomarkers, Tumor - immunology; Biosensing Techniques - methods; DNA - blood; DNA - chemistry; DNA - immunology; DNA Methylation; DNA Modification Methylases - genetics; DNA Repair Enzymes - genetics; Electrochemical Techniques - methods; Fluorescent Dyes - chemistry; Horseradish Peroxidase - chemistry; Humans; Hydrogen Peroxide - chemistry; Immunomagnetic Separation; Limit of Detection; Tumor Suppressor Proteins - genetics; 5-mC; 5-hmC; paraffin-embedded colorectal tissues; global and gene-specific DNA methylation; cancer; human serum; electrochemical platforms
• ISSN: 2379-3694; 2379-3694
• DOI: 10.1021/acssensors.8b01339

This paper reports the preparation of versatile electrochemical biosensing platforms for the simple, rapid, and PCR-independent detection of the most frequent DNA methylation marks (5-methylcytosine, 5-mC, and/or 5-hydroxymethylcytosine, 5-hmC) both at global and gene-specific levels. The implemented strategies, relying on the smart coupling of immuno-magnetic beads (MBs), specific DNA probes and amperometric detection at screen-printed carbon electrodes (SPCEs), provided sensitive and selective determination of the target methylated DNAs in less than 90 min with a great reproducibility and demonstrated feasibility for the simultaneous detection of the same or different cytosine epimarks both at global level and in different loci of the same gene or in different genes. The bioplatforms were applied to determine global methylation events in paraffin-embedded colorectal tissues and specific methylation at promoters of tumor suppressor genes in genomic DNA extracted from cancer cells and paraffin-embedded colorectal tissues, and in serum without previous DNA extraction from cancer patients.