University of Louisville
Investigating the Mechanism of AGRO100, a Novel Anti-Cancer Agent
Institution
University of Louisville
Faculty Advisor/ Mentor
Paula Bates; Simone Jueliger; Lavona Casson; Shelia Thomas
Abstract
AGRO100 is a synthetic DNA oligonucleotide that has recently entered Phase I clinical trials for the treatment of advanced cancer. AGRO100 binds to specific cellular proteins and its major target has been identified as nucleolin. However, the mechanism by which binding of AGRO100 to nucleolin inhibits cancer cell proliferation is unclear at present, in part, because the role of nucleolin in cancer biology is poorly understood. In the current study, three experiments were undertaken to further analyze the mechanism and activity of AGRO100. To elucidate the role of nucleolin in cancer cell biology, murine fibroblasts that had been transfected with human nucleolin were characterized in terms of their proliferation, invasiveness and response to chemotherapy agents. In addition, the phosphorylation status of nucleolin in untreated and AGRO100-treated cells was examined by immunoprecipitating nucleolin followed by western blotting with phosphospecific antibodies. Finally, the effect of AGRO100 treatment on NFkB signaling, which is inhibited by AGRO100 in some cell types, was assessed in prostate cancer cells. The preliminary results indicate that the nucleolin-overexpressing cells have a more aggressive phenotype than the parental cell line, that nucleolin is heavily phosphorylated but phosphorylation is not affected by AGRO100, and that AGRO100 can inhibit NFkB activity in prostate cancer cells.
Investigating the Mechanism of AGRO100, a Novel Anti-Cancer Agent
AGRO100 is a synthetic DNA oligonucleotide that has recently entered Phase I clinical trials for the treatment of advanced cancer. AGRO100 binds to specific cellular proteins and its major target has been identified as nucleolin. However, the mechanism by which binding of AGRO100 to nucleolin inhibits cancer cell proliferation is unclear at present, in part, because the role of nucleolin in cancer biology is poorly understood. In the current study, three experiments were undertaken to further analyze the mechanism and activity of AGRO100. To elucidate the role of nucleolin in cancer cell biology, murine fibroblasts that had been transfected with human nucleolin were characterized in terms of their proliferation, invasiveness and response to chemotherapy agents. In addition, the phosphorylation status of nucleolin in untreated and AGRO100-treated cells was examined by immunoprecipitating nucleolin followed by western blotting with phosphospecific antibodies. Finally, the effect of AGRO100 treatment on NFkB signaling, which is inhibited by AGRO100 in some cell types, was assessed in prostate cancer cells. The preliminary results indicate that the nucleolin-overexpressing cells have a more aggressive phenotype than the parental cell line, that nucleolin is heavily phosphorylated but phosphorylation is not affected by AGRO100, and that AGRO100 can inhibit NFkB activity in prostate cancer cells.