Anna Evans, “Effects of Short-Term Carmofur Dosing on Ceramide Levels in Cancer Cells”
Mentor: Shama Mirza, Chemistry & Biochemistry
Poster #56

Glioblastoma multiforme (GBM) is an aggressive form of brain cancer that arises in normal glial cells affecting the central nervous system. GBM is known to disrupt the regulation of acid ceramidase, the enzyme responsible for ceramide concentrations in the brain. Ceramides are signaling molecules that regulate cell pathways involved in apoptosis, growth, division, and differentiation. When ceramides are hydrolyzed by acid ceramidase, they yield a sphingosine molecule, which disrupts the signaling pathway causing cells to grow and proliferate. The chemotherapeutic drug carmofur has been shown to inhibit acid ceramidase and consequently, increase ceramide levels. In this study, U87 GBM cells were dosed with 25µM, 50µM, and 100µM concentrations of carmofur in three cycles over a period of 15 days. Consequently, each sample ended with a total concentration of 300µM over the total period. We anticipate that, carmofur will have differential increase in ceramide levels at each drug dosage regimen. Moreover, from this study, we anticipate finding a regimen that can lead to drug resistance in GBM cells. At the end of the 15-day period, ceramide extractions will be performed using a modified Folch’s method. The extracts will be analyzed by multiple reaction monitoring using a Shimadzu LCMS-8040 triple quadrupole mass spectrometer to quantify ceramide levels. In addition to ceramide analysis, GBM cells both living, and dead will be quantified and changes to morphology will be monitored. This study highlights the significance of optimizing dosing strategies for acid ceramidase inhibitors, such as carmofur, to treat GBM.