University of Louisville
A Novel Mechanism of Cancer Cell Death Triggered by G-Rich Oligonucleotides
Institution
University of Louisville
Faculty Advisor/ Mentor
Paula Bates; Shelia Thomas
Abstract
AGRO100 is a synthetic DNA oligonucleotide that has recently been tested in Phase I clinical trials for the treatment of advanced cancer. Unlike other forms of cancer treatment, AGRO100 does not appear to induce cancer cell death by triggering an apoptotic pathway. Previous research has shown that AGRO100 causes a change in the morphology of cancerous cells, which is characterized by cellular swelling, however DNA synthesis ceases. The goal of this experiment was to comparatively analyze the size difference of AGRO100 treated U937 leukemia cells and untreated U937 cells by forward-scatter flow cytometry, and to determine whether AGRO100 induces changes in protein synthesis and cell division to cause enlarged cancer cells. Cellular divisions were analyzed by reading carboxy-fluorescein diacetate, succinimidyl ester stained samples of AGRO100 treated and untreated cells by flow cytometry. 35S-methionine incorporation assay allowed for quantification of in-vitro protein synthesis, using a scintillation counting method, for AGRO100 treated and untreated U937 cells. Preliminary data indicates that AGRO100 treated cells are larger, undergo fewer cellular divisions, and synthesize more proteins than their untreated counterparts. This uncoupling of cell growth from cell division by AGRO100 treatment is the likely cause of the change in the morphology of the cancer cells, which suggests that AGRO100 may be triggering necrosis/oncosis in the cancer cells.
A Novel Mechanism of Cancer Cell Death Triggered by G-Rich Oligonucleotides
AGRO100 is a synthetic DNA oligonucleotide that has recently been tested in Phase I clinical trials for the treatment of advanced cancer. Unlike other forms of cancer treatment, AGRO100 does not appear to induce cancer cell death by triggering an apoptotic pathway. Previous research has shown that AGRO100 causes a change in the morphology of cancerous cells, which is characterized by cellular swelling, however DNA synthesis ceases. The goal of this experiment was to comparatively analyze the size difference of AGRO100 treated U937 leukemia cells and untreated U937 cells by forward-scatter flow cytometry, and to determine whether AGRO100 induces changes in protein synthesis and cell division to cause enlarged cancer cells. Cellular divisions were analyzed by reading carboxy-fluorescein diacetate, succinimidyl ester stained samples of AGRO100 treated and untreated cells by flow cytometry. 35S-methionine incorporation assay allowed for quantification of in-vitro protein synthesis, using a scintillation counting method, for AGRO100 treated and untreated U937 cells. Preliminary data indicates that AGRO100 treated cells are larger, undergo fewer cellular divisions, and synthesize more proteins than their untreated counterparts. This uncoupling of cell growth from cell division by AGRO100 treatment is the likely cause of the change in the morphology of the cancer cells, which suggests that AGRO100 may be triggering necrosis/oncosis in the cancer cells.