Combined inhibition of AXL, Lyn and p130Cas kinases block migration of triple negative breast cancer cells

Abstract

Abbreviations: AKT, RAC-a serine/threonine-protein kinase; FAK, focal adhesion kinase; EGFR, epidermal growth factor receptor; ELISA, enzyme-linked immunosorbant assay; Gas6, growth arrest specific 6; MAPK, mitogen activated protein kinases; PI3K, phosphatidylinositol 3-kinase; Pyk2, proline-rich tyrosine kinase 2; RTK, receptor tyrosine kinase; siRNA, short interfering RNA; TKI, tyrosine kinase inhibitor; TNBC, triple negative breast cancerBlocking the migration of metastatic cancer cells is a major goal in the therapy of cancer. The receptor tyrosine kinase AXL is one of the main triggers for cancer cell migration in neoplasia of breast, colon, skin, thyroid and prostate. In our study we analyzed the effect of AXL inhibition on cell motility and viability in triple negative breast cancer cell lines overexpressing AXL. Thereby we reveal that the compound BMS777607, exhibiting the lowest IC 50 values for inhibition of AXL kinase activity in the studied cell lines, attenuates cell motility to a lower extent than the kinase inhibitors MPCD84111 and SKI606. By analyzing the target kinases of MPCD84111 and SKI606 with kinase profiling assays we identified Lyn, a Src family kinase, as a target of both compounds. Knockdown of Lyn and the migrationrelated CRK-associated substrate (p130Cas), had a significant inhibitory effect on cell migration. Taken together, our findings highlight the importance of combinatorial or multikinase inhibition of non-receptor tyrosine kinases and AXL receptor tyrosine kinase in the therapy of triple negative breast cancer.

GAS6‐expressing and self‐sustaining cancer cells in 3D spheroids activate the PDK‐RSK‐mTOR pathway for survival and drug resistance

“…Therefore, we treated Caliper cells with MPCD84111, which was characterized as an efficient inhibitor for AXL‐MET‐SRC by Penzes et al . (), and sunitinib, which was characterized as a multitargeted TKI, but without the impact on MET or SRC inhibition (Penzes et al ., ). We performed ATP assays 3 days after treatment of cells with increasing concentrations of MPCD84111 or sunitinib (Fig.…”

“…Immunoprecipitation was performed as previously described (Torka, Penzes et al ., ). The home‐made clone 4G10 was used as antibody for phospho‐tyrosine precipitation in the concentration of 1 μg per 250 μg cell extract.…”

“…Cell lysis and western blot analysis were performed as previously described (Penzes et al ., ; Torka et al ., ). Primary antibodies pAKT S473 (No.…”

The miR-24-3p/p130Cas: a novel axis regulating the migration and invasion of cancer cells

“…4A). Our previous studies and others indicated that p130Cas is a proto-oncogene governing cell migration/invasion and its differential expression is related to cancer progression15162027. To test whether miR-24-3p regulates p130Cas expression, we investigated p130Cas expression in MCF7 cells after the transfection of miR-24-3p.…”

“…Here, we demonstrated that miR-24-3p is a novel regulator governing p130Cas expression. Since p130Cas plays an important role in cancer development, understanding the detailed mechanism of p130Cas regulation would provide a useful strategy for cancer therapy by targeting p130Cas2743.…”