title: A minority subpopulation of CD133(+) /EGFRvIII(+) /EGFR(-) cells acquires stemness and contributes to gefitinib resistance.
abstract: OBJECTIVE
  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.
authors.affiliation: Department of Cell Biology, Peking University Health Science Center, Beijing, China.
authors.forename: Xu-Jie
authors.initials: XJ
authors.lastname: Liu
authors.forename: Wen-Tao
authors.initials: WT
authors.lastname: Wu
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Wu
authors.forename: Feng
authors.initials: F
authors.lastname: Yin
authors.forename: Si-Hai
authors.initials: SH
authors.lastname: Ma
authors.forename: Jia-Zhen
authors.initials: JZ
authors.lastname: Qin
authors.forename: Xiu-Xiu
authors.initials: XX
authors.lastname: Liu
authors.forename: Yi-Nan
authors.initials: YN
authors.lastname: Liu
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Shuo
authors.initials: S
authors.lastname: Han
authors.forename: Kai-Yu
authors.initials: KY
authors.lastname: Liu
authors.forename: Jin-Ming
authors.initials: JM
authors.lastname: Zhang
authors.forename: Qi-Hua
authors.initials: QH
authors.lastname: He
authors.forename: Li
authors.initials: L
authors.lastname: Shen
mesh_terms: D000071916:AC133 Antigen; D000293:Adolescent; D000328:Adult; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001932:Brain Neoplasms / Q000473:pathology; D004351:Drug Resistance; D019008:Drug Resistance, Neoplasm; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D000077156:Gefitinib; D005909:Glioblastoma / Q000473:pathology; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008875:Middle Aged; D009368:Neoplasm Transplantation; D010455:Peptides / Q000378:metabolism*; D011799:Quinazolines / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000187:drug effects* / Q000502:physiology*; D055815:Young Adult
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D000970:Antineoplastic Agents; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D011799:Quinazolines; C410723:epidermal growth factor receptor VIII; C512478:EGFR protein, human; D066246:ErbB Receptors; D000077156:Gefitinib