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In situ evaluation of anticancer drug methotrexate-DNA interaction using a DNA-electrochemical biosensor and AFM
Ana Dora Rodrigues Pontinha1, Sônia Maria Alves Jorge, Ana-Maria Chiorcea Paquim
1Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Coimbra, 3004-535, Coimbra, Portugal.
Physical Chemistry Chemical Physics : PCCP
|March 2, 2011
Summary
Methotrexate (MTX) binding alters double-stranded DNA (dsDNA) structure over time. This DNA-drug interaction, visualized by atomic force microscopy, reveals structural changes and purine oxidation increases.
Area of Science:
- Electrochemistry
- Biomolecular Interactions
- Nanotechnology
Background:
- Methotrexate (MTX) is a crucial chemotherapeutic agent.
- Understanding MTX interaction with DNA is vital for drug development.
- Electrochemical biosensors offer sensitive platforms for studying biomolecular interactions.
Purpose of the Study:
- To investigate the in situ interaction between double-stranded DNA (dsDNA) and MTX.
- To characterize structural modifications of dsDNA upon MTX binding.
- To elucidate the mechanism of MTX-dsDNA interaction using electrochemical and AFM techniques.
Main Methods:
- Utilized a dsDNA-electrochemical biosensor for in situ analysis.
- Employed voltammetry to monitor electrochemical changes.
- Characterized DNA structure using atomic force microscopy (AFM) on highly oriented pyrolytic graphite (HOPG).
Main Results:
- Observed time-dependent modifications in dsDNA structure upon MTX interaction.
- AFM revealed reorganization of dsDNA network, forming denser, thicker MTX-dsDNA lattices with aggregates.
- Increased purine oxidation peaks indicated dsDNA unwinding due to MTX intercalation.
Conclusions:
- MTX binding induces significant structural changes in dsDNA.
- Electrochemical biosensors coupled with AFM provide valuable insights into drug-DNA interactions.
- The study clarifies the intercalation mechanism of MTX within dsDNA.
