Using <i>Drosophila melanogaster</i> as a Whole-Model Animal System to Elucidate the Mechanism of Action of Novel Anticancer Agents

DNA-modifying agents compose a large portion of current and phased-out anticancer agents. Interaction with DNA is linked to high levels of side effects and poor selectivity between cancerous and normal cells. This lack of specificity leads to side effects. Our lab used a prodrug strategy to design a set of novel chemotherapeutic agents. The drugs utilize a hallmark of many cancer cells, oxidative stress, for activation. This allows them to selectively target the cancerous cells, therefore limiting DNA activity in normal cells. The agents form strong electrophiles specifically when oxidized and react with DNA to form a bulky DNA lesion. A primer extension assay was carried out to visualize the DNA damage occurring on a 392-nucleotide PCR product in response to the agents. It was determined modification can occur at three out of four DNA bases. In order to test the agents and elucidate their mechanism of action, an <i>in vivo</i> model system was developed using transgenic <i>Drosophila melanogaster</i>. Mutants were generated using the Gal4-UAS system coupled with RNA interference. Knockdown was achieved for <i>Sod1</i>, a key gene in the antioxidant pathway, and <i>Ercc1</i>, a DNA repair gene. Drosophila were microinjected with the oxidatively agents, An-Hq and An-Hq₂, to determine toxicity. We compared the change in toxicity to traditional DNA modifying agents: cisplatin, chlorambucil, and 5-fluorouracil. Results of injection with the common agents were consistent with the agents’ known mechanisms of action. However, a dramatic reduction in survival was seen for both <i>Sod1</i> knockdown mutants as well as <i>Ercc1</i> mutants, indicating that our agent exhibits sensitivity to changes in both nucleotide excision repair and antioxidant capacities. The results additionally suggest that the lesions formed by our therapeutics, upon activation by elevated reactive oxygen species, will modify DNA and require nucleotide excision repair.

Name thesis:degree_name

MS

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

Arts and Sciences: Chemistry

Grantor dc:publisher

University of Cincinnati

Year dc:date

2012

Author dc:creator

• Jones, Amy R.

Contributors dc:contributor

• Merino, Edward

• Biochemistry
• Reactive Oxygen Species
• Drosophila melanogaster
• Ercc1
• DNA modifying agent
• Cancer Model
• Sod1

Statement dc:rights

• unrestricted
• This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.

Language dc:language

English

Repository record dc:identifier

http://rave.ohiolink.edu/etdc/view?acc_num=ucin1353153948

OAI identifier oai:identifier

oai:etd.ohiolink.edu:ucin1353153948