Radiosensitization and downregulation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) upon inhibition of mitogen/extracellular signal-regulated kinase (MEK) in malignant melanoma cells

Oncotarget. 2015 Jul 10;6(19):17178-91. doi: 10.18632/oncotarget.3935.

Abstract

Background: Heterogeneous nuclear ribonucleoprotein K (hnRNP K) is an important cofactor in the p53-mediated DNA damage response pathway upon ionizing radiation (IR) and exerts anti-apoptotic effects also independent of p53 pathway activation. Furthermore, hnRNP K is overexpressed in various neoplasms including malignant melanoma (MM). Here, we investigate the role of hnRNP K in the radioresistance of MM cells.

Methods and results: Our results show cytoplasmic expression of hnRNP K in human MM surgical specimens, but not in benign nevi, and a quick dose- and time-dependent upregulation in response to IR accompanied by cytoplasmic redistribution of the protein in the IPC-298 cellular tumor model carrying an activating NRAS mutation (p.Q61L). SiRNA-based knockdown of hnRNP K induced a delayed decline in γH2AX/53BP1-positive DNA repair foci upon IR. Pharmacological interference with MAPK signaling abrogated ERK phosphorylation, diminished cellular hnRNP K levels, impaired γH2AX/53BP1-foci repair and proliferative capability and increased apoptosis comparable to the observed hnRNP K knockdown phenotype in IPC-298 cells.

Conclusions: Our results indicate that pharmacological interference with MAPK signaling increases vulnerability of NRAS-mutant malignant melanoma cells to ionizing radiation along with downregulation of endogenous hnRNP K and point towards a possible use for combined MEK inhibition and localized radiation therapy of MM in the NRAS-mutant setting where BRAF inhibitors offer no clinical benefit.

Keywords: MEK inhibition; NRAS; melanoma; nRNP K; radiotherapy.

MeSH terms

  • Blotting, Western
  • DNA Repair
  • Down-Regulation
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Female
  • Flow Cytometry
  • GTP Phosphohydrolases / genetics
  • Gene Expression Regulation, Neoplastic / physiology
  • Heterogeneous-Nuclear Ribonucleoprotein K / metabolism*
  • Humans
  • Melanoma / metabolism*
  • Membrane Proteins / genetics
  • Microscopy, Fluorescence
  • Middle Aged
  • Mutation
  • RNA, Small Interfering
  • Radiation Tolerance / physiology*
  • Tissue Array Analysis
  • Transfection

Substances

  • Heterogeneous-Nuclear Ribonucleoprotein K
  • Membrane Proteins
  • RNA, Small Interfering
  • Extracellular Signal-Regulated MAP Kinases
  • GTP Phosphohydrolases
  • NRAS protein, human