We have completed mechanism of action studies on a new class of anticancer nucleosides typified by a novel nucleoside discovered in our lab, MAP-870. In order to study the mechanism of MAP-870, several experiments were completed on a colorectal adenocarcinoma cell line, HT-29, including trypan blue cell count, sulforhodamine B assays, flow cytometry of cell cycle, propidium iodide incorporation, and phosphatidylserine externalization, Caspase-Glo3/7 assays, DNA fragmentation gel, cyclophilin A release gel, PAMPA, and confocal imaging. Sulforhodamine B assays show that MAP-870 does indeed cause growth inhibition and cell death in the model tested. PAMPA assays show that MAP-870 does not appear to enter the cell via passive diffusion. Flow cytometry showed that MAP-870 doe not appear to cause cell cycle arrest or externalization of phosphatidylserine. Caspase-Glo3/7 assays demonstrated that MAP-870 does not appear to cause caspase activation. From confocal microscopy, it appears preliminarily that MAP-870 is taken up by cells, often through pseudopodia. The mechanism of MAP-870 on cancer cells must be further studied to elucidate its mechanism of action. However, preliminarily our data could point to TGFβ as a potential target pathway involving a unique, heretofore never described, cell death mechanism.
| Identifer | oai:union.ndltd.org:BGMYU2/oai:scholarsarchive.byu.edu:etd-4807 |
| Date | 02 November 2012 |
| Creators | Browning, Megan E. |
| Publisher | BYU ScholarsArchive |
| Source Sets | Brigham Young University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
| Rights | http://lib.byu.edu/about/copyright/ |
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