Identification of genes that modulate sensitivity of U373MG glioblastoma cells to cis-platinum

Anticancer Drugs. 2006 Aug;17(7):733-51. doi: 10.1097/01.cad.0000217429.67455.18.

Abstract

Scatter factor (hepatocyte growth factor) and its receptor c-Met are increasingly expressed during progression from low-grade to high-grade gliomas. Scatter factor/c-Met signaling induces glioma cell motility, invasion, angiogenesis and resistance to DNA-damaging agents. The latter is relevant to the understanding of the resistance of human gliomas to chemotherapy and radiotherapy. The goal of this study was to identify a set of genes that may contribute to scatter factor-mediated protection of U373MG cells against cis-platinum, a DNA cross-linking agent. We used DNA microarray assays, confirmatory semiquantitative reverse transcription-polymerase chain reaction analysis and functional assays to identify genes involved in the scatter factor-induced resistance of U373MG to cis-platinum. We identified a group of genes that are overexpressed in cells treated with scatter factor plus cis-platinum relative to cells treated with cis-platinum alone and confirmed some of these gene expression alterations by reverse transcription-polymerase chain reaction. Inhibiting the expression of three of these genes--polycystic kidney disease 1, amplified in breast cancer 1 and DEAD/H box helicase 21--using small interfering RNAs reduced survival of cis-platinum-treated cells and partially reversed the scatter factor protection against cis-platinum. Dominant-negative Akt and IkappaB super-repressor expression vectors inhibited the scatter factor protection, and abrogated the ability of scatter factor to alter the expression of DEAD/H box helicase 21 and polycystin (PKD1) within the context of cis-platinum exposure. The Akt and nuclear factor-kappaB inhibitors had no effect on amplified in breast cancer 1 expression. These studies implicate DEAD/H box helicase 21, polycystin (PKD1) and amplified in breast cancer 1 as novel transcription-dependent regulators of scatter factor-mediated glioma cell protection against cytotoxic death, and identify other potential regulators for future study.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antineoplastic Agents / therapeutic use*
  • Blotting, Western
  • Brain Neoplasms / drug therapy*
  • Brain Neoplasms / genetics*
  • Cell Line, Tumor
  • Cisplatin / therapeutic use*
  • DNA Fingerprinting
  • Down-Regulation / drug effects
  • Drug Resistance, Neoplasm
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genetic Vectors
  • Glioblastoma / drug therapy*
  • Glioblastoma / genetics*
  • Hepatocyte Growth Factor / physiology*
  • Humans
  • NF-kappa B / genetics
  • Oligonucleotide Array Sequence Analysis
  • Oncogene Protein v-akt / genetics
  • Proto-Oncogene Proteins c-met / physiology*
  • RNA, Neoplasm / isolation & purification
  • RNA, Small Interfering
  • Transfection

Substances

  • Antineoplastic Agents
  • NF-kappa B
  • RNA, Neoplasm
  • RNA, Small Interfering
  • Hepatocyte Growth Factor
  • Proto-Oncogene Proteins c-met
  • Oncogene Protein v-akt
  • Cisplatin