| Autor: |
Gurgis F; School of Medical Sciences, Department of Pharmacology, The University of Sydney , Sydney, NSW 2006, Australia., Åkerfeldt MC; School of Medical Sciences, Department of Pharmacology, The University of Sydney , Sydney, NSW 2006, Australia., Heng B; Faculty of Medicine and Health Sciences, Macquarie University , NSW 2109, Australia., Wong C; Children's Medical Research Institute, The University of Sydney , Sydney, NSW 2145, Australia., Adams S; Faculty of Medicine and Health Sciences, Macquarie University , NSW 2109, Australia., Guillemin GJ; Faculty of Medicine and Health Sciences, Macquarie University , NSW 2109, Australia., Johns TG; MIMR-PHI Institute of Medical Research, 27-31 Wright Street, Clayton, VIC 3168, Australia; Monash University, Wellington Road, Clayton, VIC 3800, Australia., Chircop M; Children's Medical Research Institute, The University of Sydney , Sydney, NSW 2145, Australia., Munoz L; School of Medical Sciences, Department of Pharmacology, The University of Sydney , Sydney, NSW 2006, Australia. |
| Abstrakt: |
MAPK-activated protein kinase 2 (MK2) is a checkpoint kinase involved in the DNA damage response. MK2 inhibition enhances the efficacy of chemotherapeutic agents; however, whether MK2 inhibition alone, without concurrent chemotherapy, would attenuate survival of cancer cells has not been investigated. CMPD1 is a widely used non-ATP competitive inhibitor that prevents MK2 phosphorylation. We employed CMPD1 together with MK2 knock-down and ATP-competitive MK2 inhibitor III (MK2i) in a panel of glioblastoma cells to assess whether MK2 inhibition could induce cancer cell death. While CMPD1 was effective at selective killing of cancer cells, MK2i and MK2 knock-down had no effect on viability of glioblastoma cells. CMPD1 treatment induced a significant G2/M arrest but MK2i-treated cells were only minimally arrested at G1 phase. Intriguingly, at doses that were cytotoxic to glioblastoma cells, CMPD1 did not inhibit phosphorylation of MK2 and of its downstream substrate Hsp27. These results suggest that CMPD1 exhibits cytotoxic activity independently of MK2 inhibition. Indeed, we identified tubulin as a primary target of the CMPD1 cytotoxic activity. This study demonstrates how functional and mechanistic studies with appropriate selection of test compounds, combining genetic knock-down and pharmacological inhibition, coordinating timing and dose levels enabled us to uncover the primary target of an MK2 inhibitor commonly used in the research community. Tubulin is emerging as one of the most common non-kinase targets for kinase inhibitors and we propose that potential tubulin-targeting activity should be assessed in preclinical pharmacology studies of all novel kinase inhibitors. |
