Study on the interaction between doxorubicin and Deoxyribonucleic acid with the use of methylene blue as a probe

Abstract

Neste trabalho, a interação de doxorrubicina com DNA (obtido de timo de bezerro) em fita dupla foi investigada através de técnicas de espectrofotometria UV-Vis, voltametria e espectrofluorometria, usando azul de metrileno (MB) como marcador. O comportamento voltamétrico da doxorrubicina foi investigado em eletrodo de carbono vítreo usando voltametria de pulso diferencial. A doxorrubicina é reduzida, produzindo um pico de redução. Os dois estudos, espectrofotometria UV-Vis e voltametria de pulso diferencial, confirmam a reação de intercalação. Os resultados mostraram que a doxorrubicina e a molécula de MB foram intercaladas na dupla hélice do DNA. A constante de ligação aparente de doxorrubicina com DNA foi 3,2 × 10 4 L mol -1 . O sinal de fluorescência da doxorrubicina e azul de metileno é suprimido com a adição de DNA. A equação de Stern-Volmer baseou-se na supressão do sinal de fluorescência da doxorrubicina.In this work, the interaction of doxorubicin with calf thymus double strand Deoxyribonucleic acid (ds-DNA) has been investigated with the use of Methylene Blue (MB) dye as a probe by the application of UV-Vis spectrophotometry, voltammetry and spectrofluorometry. The voltammetric behavior of doxorubicin has been investigated at glassy carbon electrode using differential pulse voltammetry. Doxorubicin is reduced, yielding one reduction peak. Both UV-Vis spectrophotometry and differential pulse voltammetry studies confirm the intercalation reaction. The results showed that both doxorubicin and the MB molecule could intercalate into the double helix of the DNA. The apparent binding constant of doxorubicin with DNA has been found to be 3.2 × 10 4 L mol -1 . The fluorescence signal of doxorubicin and methylene blue was quenched with DNA addition. The Stern-Volmer equation was plotted based on quenching fluorescence signal of doxorubicin.Keywords: doxorubicin, DNA, chemotherapy, spectrophotometry, voltammetry, spectrofluorometry IntroductionStudy of interactions between drugs and DNA is very interesting and significant not only in understanding the mechanism of interaction, but also for the design of new drugs. 1,2 However mechanism of interactions between drug molecules and DNA is still relatively little known. It is necessary to introduce more simple methods for investigating the mechanism of interaction. By understanding the mechanism of interaction, designing of new DNA-targeted drugs and the screening of these in vitro will be possible.A great variety of substances, including several agents of importance in cancer chemotherapy, 3 are known to bind to DNA by intercalation. 4 Attention has been concentrated on the classical intercalating drugs, acridines and ethidium bromide. [4][5][6] Studies on the binding of various dyes, drugs and antibiotics to DNA and chromatin have contributed to the understanding of the structure of these macromolecules, 6-14 and have suggested possible mechanisms of the biological activity of some drugs. 3 Molecular models of the intercalation of some drugs into DNA have been describ...

Exploration of Electrochemical Intermediates of the Anticancer Drug Doxorubicin Hydrochloride Using Cyclic Voltammetry and Simulation Studies with an Evaluation for Its Interaction with DNA

“…The apparent binding constant obtained from double reciprocal plot (see (6), Figure 5) was (1.66 ± 0.12) × 10 4 M −1 (Table 2) indicating that there was excellent agreement between the two approaches. A previous study by Hajian and coworkers [42] showed apparent binding constant for the interaction of doxorubicin hydrochloride-CT DNA in phosphate buffer at pH 7.4 as agreement with our results. The slight deviation between the previous and present results may be due to different types of measurements and different reaction media in the two studies.…”

DNA-binding studies of complex of Pt(bpy)(pip)]2+ and [Pt(bpy)(hpip)]2+ by electrochemical methods: development of an electrochemical DNA biosensor

“…Because it is well established that if there is a shift towards to negative potential, interaction between DNA and regarding complex is mainly electrostatic since electrostatic interaction between DNA and the complexes makes reduction at the surface more difficult. However, in this study, the peak potential appears at the more positive values in the presence of DNA, indicating an intercalation binding mode [32][33][34][35][36][37][38]. Owing to the fact that 1 and 2 possess positive charge, it is intrinsically expected that both platinum complexes have a binding affinity to dsDNA by an electrostatic interaction as previously reported for other positively charged molecules.…”

Application of High‐Content Analysis in Clinical Cytology for Translational Safety Biomarkers of Drug‐Induced Toxicity for Lymphoma Chemotherapy

“…It may be attributable to a compensatory adaptive response to chemical stress that is overwhelmed as the stress progresses; this has shown to be the case, for example, with diquat [11] and may well be the case for Mz and AC. The anthracyclines are known to produce anticancer effects by their intercalation into DNA to inhibit the cell cycle [12]. This is associated with nuclear swelling.…”