MELK-T1, a small-molecule inhibitor of protein kinase MELK, decreases DNA-damage tolerance in proliferating cancer cells

Biosci Rep. 2015 Oct 2;35(6):e00267. doi: 10.1042/BSR20150194.

Abstract

Maternal embryonic leucine zipper kinase (MELK), a serine/threonine protein kinase, has oncogenic properties and is overexpressed in many cancer cells. The oncogenic function of MELK is attributed to its capacity to disable critical cell-cycle checkpoints and reduce replication stress. Most functional studies have relied on the use of siRNA/shRNA-mediated gene silencing. In the present study, we have explored the biological function of MELK using MELK-T1, a novel and selective small-molecule inhibitor. Strikingly, MELK-T1 triggered a rapid and proteasome-dependent degradation of the MELK protein. Treatment of MCF-7 (Michigan Cancer Foundation-7) breast adenocarcinoma cells with MELK-T1 induced the accumulation of stalled replication forks and double-strand breaks that culminated in a replicative senescence phenotype. This phenotype correlated with a rapid and long-lasting ataxia telangiectasia-mutated (ATM) activation and phosphorylation of checkpoint kinase 2 (CHK2). Furthermore, MELK-T1 induced a strong phosphorylation of p53 (cellular tumour antigen p53), a prolonged up-regulation of p21 (cyclin-dependent kinase inhibitor 1) and a down-regulation of FOXM1 (Forkhead Box M1) target genes. Our data indicate that MELK is a key stimulator of proliferation by its ability to increase the threshold for DNA-damage tolerance (DDT). Thus, targeting MELK by the inhibition of both its catalytic activity and its protein stability might sensitize tumours to DNA-damaging agents or radiation therapy by lowering the DNA-damage threshold.

Keywords: chemical biology; deoxyribonucleic acid (DNA) damage response; maternal embryonic leucine zipper kinase (MELK) kinase; senescence; small molecule inhibitors.

MeSH terms

  • Apoptosis / drug effects
  • Ataxia Telangiectasia Mutated Proteins / biosynthesis
  • Azepines / administration & dosage*
  • Benzamides / administration & dosage*
  • Breast Neoplasms / drug therapy
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Breast Neoplasms / radiotherapy
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • DNA Damage / drug effects*
  • Enzyme Inhibitors / administration & dosage*
  • Female
  • Forkhead Box Protein M1
  • Forkhead Transcription Factors / biosynthesis
  • Gene Expression Regulation, Neoplastic
  • Humans
  • MCF-7 Cells
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / biosynthesis*
  • Protein Serine-Threonine Kinases / genetics

Substances

  • Azepines
  • Benzamides
  • Enzyme Inhibitors
  • FOXM1 protein, human
  • Forkhead Box Protein M1
  • Forkhead Transcription Factors
  • MELK-T1
  • MELK protein, human
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • Protein Serine-Threonine Kinases