Date of Graduation

Summer 2008

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Richard Garrad

Abstract

Extracellular ATP has the capability to induce a variety of physiological effects via the interaction with P2 receptors. Recent studies suggest that ATP has the potential to induce apoptosis in certain types of colon, pancreatic, and breast cancer. We have attempted to confirm that ATP does induce apoptosis in an estrogen-independent breast cancer cell line, MDA-MB-231. In addition, we tested certain ATP derivatives, ATPγS and 2MeSATP, for the ability to induce apoptosis compared to that of ATP. Other studies conducted in the lab suggested that the P2Y1 receptor is most prevalent on this cell line. This receptor has a greater response for 2MeSATP and lower for ATPγS compared to ATP. We used fluorescence microscopy, western blot, and trypan blue exclusion to determine the amount of induction. Antibodies indicated cleaved caspase-7, -9, and PARP were at the greatest quantities using the 2MeSATP as a P2 receptor agonist, with the largest amount of cleavage at 10-4 M. Additionally, we studied the effect on this induction in the presence of CaCl2, CaPO4 using the same methods. Treatment with nucleotide agonists in the presence of calcium lowered the activation of caspase-7, -9, and PARP, however decreased the amount of viable cells in solution.

Keywords

Purinergic receptor, Breast Cancer, ATP, MDA-MB-231, Apoptosis

Subject Categories

Medical Molecular Biology

Copyright

© Caleb Masterson

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