GW525701 Treatment Alters Cellular Morphology and Decreases Triple-Negative Breast Cancer Viability




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Purpose: Breast cancer is the second leading cause of death in women in the United States. Triple-negative breast cancer (TNBC) is named for its lack of estrogen (ER) and progesterone (PR) receptors, as well as HER2. The lack of receptors makes these tumors increasingly more difficult to treat, resulting in worser clinical outcomes compared to other receptor-positive subtypes. Kinase inhibitor compounds have been of great interest for treating TNBC patients. Specific kinase inhibitors have been successful against other cancer subtypes, namely chronic myeloid leukemia (CML), which has substantially bettered the outcomes associated with this disease. However, in total, only 72 kinase inhibitors have been approved for clinical use, many of which are not used as cancer therapeutics at all, let alone for breast cancer. Much more research remains to determine the role of other, understudied kinases in cancer proliferation and survival. One kinase inhibitor compound, GW525701, targets STK10, TNIK, SLK, MAP4K4, MINK, and DDR1, none of which are targeted by FDA-approved therapeutics. If treating cancer cells with this kinase inhibitor compound reduces cancer survival and proliferation, then it may be a potential future therapeutic, and will give better insight into the mechanisms controlling TNBC survival. Methods: Breast cancer cells (4IC, BT-549, MCF-7) were treated with GW525701 for 72 hours at a range of concentrations (1nM, 10nM, 100nM, 1µM, 10µM), then stained with phalloidin to visualize cytoskeletal changes. Cellular proliferation and viability were tested using a Cell Titer Glo assay. Migration changes were visualized and quantified using a Boyden migration chamber and ImageJ analysis. All treatment groups were compared to vehicle-treated controls. Results:GW525701 treatment at all concentrations caused morphologic changes consistent with senescence and apoptotic cell death. Cellular proliferation and viability were inversely correlated with GW525701 treatment concentration. Conclusions: Treatment with GW525701 may give insight into the roles of STK10, TNIK, SLK, MAP4K4, MINK, and DDR1 in TNBC survival. This information will be crucial to determining an effective means of targeting TNBC in a therapeutic manner in the future. The specific kinase(s) responsible for the phenotypic outcomes in this experiment will be elucidated in the future as well.