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Neuro-Oncology 2000 2(3):159-163; doi:10.1093/neuonc/2.3.159
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© 2000 by the Society forNeuro-Oncology

Analysis of genomic rearrangements associated with EGFRvIII expressionsuggests involvement of Alu repeat elements

Lori Frederick, Greg Eley, Xiao-Yang Wang and C. David James2

Department of Laboratory Medicine and Pathology and Tumor BiologyProgram, Mayo Clinic and Foundation, Rochester, MN 55905

2 Address correspondence and reprint requests to C. David James, Mayo Clinic,200 First St., S.W., Hilton Bldg. Rm. 820-D, Rochester, MN 55905.


  Abstract

We have developed a polymerase chain reaction (PCR)-based strategy for thesynthesis and analysis of rearranged epidermal growth factor receptor(EGFR) fragments associated with the vIII mutant receptor expressedin glioblastomas with EGFR amplification. The sequencing of aberranttumor fragments showed that intragenic deletion rearrangements consistentlyinvolve an approximately 600-bp region in intron 7 of EGFR andseveral rearrangement sites interspersed throughout the large (>100 kb)first intron of this gene. Examination of the intron 7 breakpoint regionrevealed an Alu repeat element, and all intron 7 rearrangement sites werelocated within or downstream of this repeat sequence. Analysis of intron 1 forsimilar sequences resulted in the identification of 11 sites containing>80% homology with parts of the Alu element in intron 7. Reversetranscriptase-PCR and/or Western analysis of the tumors showed the presence ofEGFRvIII cDNAs and/or proteins, respectively, in all cases for which arearranged genomic fragment was generated by long-range PCR. Collectively,these data suggest that EGFR rearrangements, associated with thesynthesis of the most common EGFR mutant, are mediated by a specific sequenceelement.

Received March 8, 2000; Accepted March 30, 2000


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