Alexis Quintero

Helios Scholar

Metastatic breast cancer affects 30% of women who develop breast cancer. It has been shown that up to 34% of breast tumors have a deficiency of the tumor suppressor gene Inhibitor of Growth 4 (ING4). It has been shown that migration of ING4-deleted triple negative breast cancer (TNBC) cells was induced by C-X-C motif chemokine ligand 10 (Cxcl10). Signaling for migration required the C-X-C motif chemokine receptor 3 (Cxcr3) and the epidermal growth factor receptor (Egfr), which was determined using receptor inhibitor. To determine whether Cxcl10 induced cell migration in the absence of the CXCR3 or EGFR genes, ING4-deleted MDAmb231 TNBC cells were genetically modified using CRISPR-Cas9 to delete CXCR3 or EGFR. Cxcl10-induced cell migration was evaluated using transwell migration assays. The results showed that Cxcl10 did not induce migration in the CXCR3-deleted or EGFR-deleted cells, indicating that Cxcr3 and Egfr were required for Cxcl10-induced migration. However, as Cxcl10 did not induce migration in the vector control cells, the experiment will be repeated to determine the Cxcr3 and Egfr requirements. To determine Cxcr3 or Egfr presence in the knockout cell lines, we used immunofluorescent staining to visualize the receptors. The results showed that Cxcr3 and Egfr were present in the vector control and CXCR3 or EGFR-deleted cells, indicating that the receptors may not be effectively deleted from these cells. By better understanding the requirements of Cxcr3 and Egfr in Cxcl10-induced cell migration, innovative targeted therapies can be developed to improve ING4-deficient metastatic breast cancer patient survival.