Distinct patterns of alteration of myc genes associated with integration of human papillomavirus type 16 or type 45 DNA in two genital tumours

Unité Mixte Institut Pasteur/INSERM (U.190), Institut Pasteur, 25 rue du Docteur Roux, F-75724 Paris cedex 15, France 1 Laboratoire de Pathologie, Section Médicale et Hospitalière, Institut Curie, Paris, France 2 Author for correspondence: Gérard Orth. Fax +33 1 45 68 89 66. e-mail gorth{at}pasteur.fr We previously described two genital carcinomas (IC2, IC4) containing human papillomavirus type 16 (HPV-16)- or HPV-18-related sequences integrated in chromosomal bands containing the c- myc (8q24) or N- myc (2p24) gene, respectively. The c- myc gene was rearranged and amplified in IC2 cells without evidence of overexpression. The N- myc gene was amplified and highly transcribed in IC4 cells. Here, the sequence of an 8039 bp IC4 DNA fragment containing the integrated viral sequences and the cellular junctions is reported. A 3948 bp segment of the genome of HPV-45 encompassing the upstream regulatory region and the E6 and E7 ORFs was integrated into the untranslated part of N- myc exon 3, upstream of the N- myc polyadenylation signal. Both N- myc and HPV-45 sequences were amplified 10- to 20-fold. The 3' ends of the major N- myc transcript were mapped upstream of the 5' junction. A minor N- myc /HPV-45 fusion transcript was also identified, as well as two abundant transcripts from the HPV-45 E6–E7 region. Large amounts of N-myc protein were detected in IC4 cells. A major alteration of c- myc sequences in IC2 cells involved the insertion of a non-coding sequence into the second intron and their co-amplification with the third exon, without any evidence for the integration of HPV-16 sequences within or close to the gene. Different patterns of myc gene alterations may thus be associated with integration of HPV DNA in genital tumours, including the activation of the protooncogene via a mechanism of insertional mutagenesis and/or gene amplification.