Publication Date

2016-05-09

Document Type

Honors Thesis

Department

Biochemistry

Keywords

Nuclear magnetic resonance, Cisplatin, DNA replication, DNA-Analysis, Cisplatin-Effect of temperature on, NMR, DNA, Base pair opening, Melting studies

Abstract

Cisplatin is a platinum-based anti-cancer drug that is used to treat a variety of cancers; although it is best known for its effectiveness in treating testicular cancer. Since its serendipitous discovery and subsequent FDA approval in 1978, 1 cisplatin’s mechanism of action has been widely studied. Cisplatin is able to enter the cell through passive diffusion and hydrolyze to a more active form. It then forms 1,2-intrastrand cross-links, distorting the helical structure of DNA. While the distorted helical structure of DNA may be enough to stop replication and transcription, leading to cell death, other mechanisms have also been proposed.2 The purpose of my research is to further the mechanistic studies of cisplatin-modified DNA by using a DNA 9-mer. A 9-mer control duplex and cisplatin-modified 9-mer were first prepared and HPLC purified. After method development using an 11-mer duplex, analysis of the 9-mer duplexes was performed. 1 H NMR was the primary analysis component, with experiments including melting experiments, 2D NOESY and COSY experiments and base pair opening experiments. It was found that the cisplatin-modified duplex is less stable than the control by about 10°C, which is not surprising given the nature by which platinum disrupts the helical structure of DNA. Base pair opening experiments were not performed on the modified duplex, however base pair opening experiments on the control duplex were performed, and the data were collected for future analysis.

Language

English

Comments

89 pages : illustrations (some color). Honors project, Smith College, 2016. Includes bibliographical references (pages 87-89)

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