Evaluation of the Cytotoxic Profile of Metformin and Y15 in Platinum Resistance Ovarian Cancer Cells
Abstract
Metformin, an antidiabetic drug, has been previously shown to induce cytotoxicity in platinum resistant ovarian cancer cells and increase phosphorylation of Focal Adhesion Kinase (FAK), a tyrosine kinase implicated in the development of this platinum resistance. Therefore, we evaluated the combined cytotoxic efficacy of Metformin and the focal adhesion kinase inhibitor 1,2,4,5-Benzene Tetra amine tetra hydrochloride (Y15) in platinum resistant OVCAR 3 ovarian cancer cells. The cells were treated with concentrations of Y15 and Metformin to determine IC50 values, which were then used to treat cells with Y15 and Metformin separately and in combination. DNA fragmentation and poly ADP ribose polymerase (PARP) cleavage assays were performed to evaluate mechanisms of cell death. We further utilized a western blot to evaluate the expression of phosphorylated FAK, p53, p21, and pAKT in response to the treatments. The results showed that in combination, Y15 significantly increased efficacy of metformin compared to the metformin only treatment. Cell death by apoptosis was confirmed by PARP cleavage and presence of DNA fragments in treatment groups. The combination treatment of metformin and Y15 significantly downregulated phosphorylated FAK expression, confirming reduced FAK activity. This reduced FAK auto phosphorylation correlated with increased p53 and p21 expression. Therefore, Y15 significantly enhances the cytotoxic profile of metformin in platinum resistant OVCAR 3 cells. This study is the first to report a FAK dependent cytotoxic mechanism of metformin in ovarian cancer, leading to the understanding of the cooperation between metformin and Y15 to inhibit FAK activity in platinum resistant ovarian cancer cells.
Faculty Sponsors
Dr. Deanne Roopnarine, Dr. Julie Torruellas Garcia, Dr. Robert Smith
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-8-2016 1:00 PM
End Date
4-8-2016 5:30 PM
Evaluation of the Cytotoxic Profile of Metformin and Y15 in Platinum Resistance Ovarian Cancer Cells
Alvin Sherman Library
Metformin, an antidiabetic drug, has been previously shown to induce cytotoxicity in platinum resistant ovarian cancer cells and increase phosphorylation of Focal Adhesion Kinase (FAK), a tyrosine kinase implicated in the development of this platinum resistance. Therefore, we evaluated the combined cytotoxic efficacy of Metformin and the focal adhesion kinase inhibitor 1,2,4,5-Benzene Tetra amine tetra hydrochloride (Y15) in platinum resistant OVCAR 3 ovarian cancer cells. The cells were treated with concentrations of Y15 and Metformin to determine IC50 values, which were then used to treat cells with Y15 and Metformin separately and in combination. DNA fragmentation and poly ADP ribose polymerase (PARP) cleavage assays were performed to evaluate mechanisms of cell death. We further utilized a western blot to evaluate the expression of phosphorylated FAK, p53, p21, and pAKT in response to the treatments. The results showed that in combination, Y15 significantly increased efficacy of metformin compared to the metformin only treatment. Cell death by apoptosis was confirmed by PARP cleavage and presence of DNA fragments in treatment groups. The combination treatment of metformin and Y15 significantly downregulated phosphorylated FAK expression, confirming reduced FAK activity. This reduced FAK auto phosphorylation correlated with increased p53 and p21 expression. Therefore, Y15 significantly enhances the cytotoxic profile of metformin in platinum resistant OVCAR 3 cells. This study is the first to report a FAK dependent cytotoxic mechanism of metformin in ovarian cancer, leading to the understanding of the cooperation between metformin and Y15 to inhibit FAK activity in platinum resistant ovarian cancer cells.
