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.

Abstract

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.

• Authors: Eloy Povedano, Víctor Ruiz-Valdepeñas Montiel, Alejandro Valverde, Fernando Navarro-Villoslada, Paloma Yáñez-Sedeño, María Pedrero, Ana Montero-Calle, Rodrigo Barderas, Alberto Peláez-García, Marta Mendiola, David Hardisson, Jaime Feliú, Jordi Camps, Elisabet Rodríguez-Tomàs, Jorge Joven, Meritxell Arenas, Susana Campuzano, José M Pingarrón
• Journal: ACS sensors (ACS Sens) Vol. 4 Issue 1 Pg. 227-234 (01 25 2019) ISSN: 2379-3694 [Electronic] United States
• PMID: 30499292 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Biomarkers, Tumor
• Fluorescent Dyes
• RASSF1 protein, human
• Tumor Suppressor Proteins
• 5-hydroxymethylcytosine
• 5-Methylcytosine
• DNA
• Hydrogen Peroxide
• Horseradish Peroxidase
• DNA Modification Methylases
• MGMT protein, human
• DNA Repair Enzymes

Topics

• 5-Methylcytosine (analogs & derivatives, blood, immunology)
• Antibodies, Monoclonal (immunology)
• Armoracia (enzymology)
• Biomarkers, Tumor (blood, chemistry, immunology)
• Biosensing Techniques (methods)
• DNA (blood, chemistry, 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)