Chemo-sensitisation of HeLa cells to etoposide by a benzoxazine in the absence of DNA-PK inhibition.

Abstract

The benzoaxines have been developed from structurally similar chromones as specific inhibitors of the PI3K family to sensitize cancer cells to the effects of chemotherapeutic agents; most have been shown to do this through specific inhibition of DNA-PK and DNA repair mechanisms. In this study we examined the benzoxazine, 2-((3-methoxybut-3en-2-yl)amino)-8methyl-4H-benzo[1,3]oxazin-4one (LTUSI54). This compound had no DNA-PK or PI3K inhibitory activity but still sensitized HeLa cells to the effects of Etoposide. LTUSI54 works synergistically with Etoposide to inhibit growth of HeLa cells and sub G1 analysis indicates that this is not due to an increase in apoptosis. LTUSI54 neither enhances DSB formation due to Etoposide nor does it delay the repair of such damage. Cell cycle analysis shows a clear G2 block with Etoposide alone while, in combination with LTUSI54 there is an additional S phase arrest. Phospho-kinase analysis indicated that LTUSI54 engages key regulators of cell cycle progression, specifically p38α, p53 and ERK 1/2. From our results we hypothesize that LTUSI54 is promoting the cell cycle arrest through activation of p38α pathways, independent of p53 mechanisms. This results in a decrease in p53 phosphorylation and hence, restricted apoptosis. Changes in cell number appear to be the result of p38α pathways disrupting cell cycle progression, at the S and G2 checkpoints. Further investigation into the finer mechanisms by which LTUSI54 effects cell cycle progression would be of great interest in assessing this compound as a chemosensitising agent.

• Authors: Cheree Fitzgibbon, Saleh Ihmaid, Jasim Al-Rawi, Terri Meehan-Andrews, Christopher Bradley
• Journal: Investigational new drugs (Invest New Drugs) Vol. 31 Issue 6 Pg. 1466-75 (Dec 2013) ISSN: 1573-0646 [Electronic] United States
• PMID: 24057508 (Publication Type: Journal Article)

Chemical References

• 2-((3-methoxybut-3-en-2-yl)amino)-8-methyl-4H-benzo(1,3)oxazin-4-one
• Antineoplastic Agents
• Benzoxazines
• H2AX protein, human
• Histones
• Topoisomerase II Inhibitors
• Etoposide
• DNA-Activated Protein Kinase
• MAPK1 protein, human
• Mitogen-Activated Protein Kinase 1
• Mitogen-Activated Protein Kinase 3
• Alanine

Topics

• Alanine (analogs & derivatives, pharmacology)
• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Benzoxazines (pharmacology)
• Cell Cycle (drug effects)
• Cell Proliferation (drug effects)
• Cell Survival (drug effects)
• DNA-Activated Protein Kinase (metabolism)
• Etoposide (pharmacology)
• HeLa Cells
• Histones (metabolism)
• Humans
• Mitogen-Activated Protein Kinase 1 (metabolism)
• Mitogen-Activated Protein Kinase 3 (metabolism)
• Topoisomerase II Inhibitors (pharmacology)