A minority subpopulation of CD133(+) /EGFRvIII(+) /EGFR(-) cells acquires stemness and contributes to gefitinib resistance

CNS Neurosci Ther. 2013 Jul;19(7):494-502. doi: 10.1111/cns.12092. Epub 2013 Apr 10.

Abstract

Aims: To study the contribution of epidermal growth factor receptor variant III (EGFRvIII) to glioblastoma multiforme (GBM) stemness and gefitinib resistance.

Methods: CD133(+) and CD133(-) cells were separated from EGFRvIII(+) clinical specimens of three patients with newly diagnosed GBM. Then, RT-PCR was performed to evaluate EGFRvIII and EGFR expression in CD133(+) and CD133(-) cells. The tumorigenicity and stemness of CD133(+) cells was verified by intracranial implantation of 5 × 10(3) cells into immunodeficient NOD/SCID mice. Finally, cells were evaluated for their sensitivity to EGFR tyrosine kinase inhibition by gefitinib.

Results: RT-PCR results showed that the sorted CD133(+) cells expressed EGFRvIII exclusively, while the CD133(-) cells expressed both EGFRvIII and EGFR. At 6-8 weeks postimplantation, CD133(+) /EGFRvIII(+) /EGFR(-) cells formed intracranial tumors. Cell counting kit-8 results showed that the IC50 values of the three isolated EGFRvIII(+) cell lines treated with gefitinib were 14.44, 16.00, and 14.66 μM, respectively, whereas the IC50 value of an isolated EGFRvIII(-) cell line was 8.57 μM.

Conclusions: EGFRvIII contributes to the stemness of cancer stem cells through coexpression with CD133 in GBMs. Furthermore, CD133(+) /EGFRvIII(+) /EGFR(-) cells have the ability to initiate tumor formation and may contribute to gefitinib resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen
  • Adolescent
  • Adult
  • Animals
  • Antigens, CD / metabolism*
  • Antineoplastic Agents / pharmacology*
  • Brain Neoplasms / pathology
  • Drug Resistance
  • Drug Resistance, Neoplasm
  • ErbB Receptors / antagonists & inhibitors
  • ErbB Receptors / metabolism*
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gefitinib
  • Glioblastoma / pathology
  • Glycoproteins / metabolism*
  • Humans
  • Immunohistochemistry
  • Immunomagnetic Separation
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Middle Aged
  • Neoplasm Transplantation
  • Peptides / metabolism*
  • Quinazolines / pharmacology*
  • Real-Time Polymerase Chain Reaction
  • Stem Cells / drug effects*
  • Stem Cells / physiology*
  • Young Adult

Substances

  • AC133 Antigen
  • Antigens, CD
  • Antineoplastic Agents
  • Glycoproteins
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse
  • Quinazolines
  • epidermal growth factor receptor VIII
  • EGFR protein, human
  • ErbB Receptors
  • Gefitinib