Epidermal growth factor receptor mutation type III transfected into a small cell lung cancer cell line is predominantly localized at the cell surface and enhances the malignant phenotype.
In the present study we transfected the epidermal growth factor receptor (EGFR)-negative small cell lung cancer cell line, GLC3, with the type III EGFR mutation (EGFRvIII). The EGFRvIII protein could be detected by Western blot analysis as a 145-kDa protein, which by immunohistochemistry appeared to be localized at the cell surface. Ultrastructurally EGFRvIII was expressed mainly at the cell surface with clusters at cell-cell contacts. In the in vitro invasion assay, GLC3-EGFRvIII cells had a approximately 5-fold increased invasion compared with uninduced GLC3-EGFRvIII, GLC3-Tet-On and the parental cell line. GLC3-Tet-On appeared uniform in size with adherence junctions at cell-cell contacts. In uninduced GLC3-EGFRvIII cells adherence junctions were also present but less distinct. In doxycycline-pretreated GLC3-EGFRvIII cells, adherence junctions were absent. We conclude that the expression of EGFRvIII results in a more malignant phenotype. This effect appears to involve the disruption of adherence junctions.
Damstrup
Lars
L
Department of Radiation Biology, The Finsen Center, University Hospital Copenhagen, Copenhagen, Denmark.
Wandahl Pedersen
Mikkel
M
Bastholm
Lone
L
Elling
Folmer
F
Skovgaard Poulsen
Hans
H
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015153:Blotting, Western; D018288:Carcinoma, Small Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002450:Cell Communication; D003094:Collagen / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004318:Doxycycline / Q000494:pharmacology; D004338:Drug Combinations; D066246:ErbB Receptors / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques; D007797:Laminin / Q000737:chemistry; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008854:Microscopy, Electron; D009154:Mutation; D009361:Neoplasm Invasiveness; D011509:Proteoglycans / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014407:Tumor Cells, Cultured
D000900:Anti-Bacterial Agents; D017931:DNA Primers; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012333:RNA, Messenger; C410723:epidermal growth factor receptor VIII; C059485:matrigel; D003094:Collagen; D066246:ErbB Receptors; D004318:Doxycycline