A comprehensive analysis of HPV integration loci in anogenital lesions combining transcript and genome-based amplification techniques.
Persistent infections with high-risk human papillomaviruses (HPVs) induce dysplastic lesions of the lower genital tract. Some of these lesions eventually progress to invasive cancers, particularly of the uterine cervix. In many advanced preneoplastic cervical lesions and most derived carcinomas, HPV genomes are found to be integrated into the host cell chromosomes. Although HPV integration seems to play an important role in the progression of cervical dysplasia, the underlying mechanisms are still unclear. To investigate the pathogenic role of genomic integration of HPV genomes in greater detail, we analysed integration sites of HPV16 and 18 genomes in 21 anogenital precancerous and cancerous lesions using a ligation-mediated chain reaction (DIPS) and the recently described amplification of papilloma virus oncogene transcripts (APOT) assay. On the genomic level, only singular integration events were observed in individual neoplastic cell clones. At many integration sites, a short overlap between HPV and genomic sequences was observed, suggesting that the integration of HPV genomes is mediated by nonhomologous sequence-specific recombination. APOT analysis revealed that the majority of integrated HPV genomes was actively transcribed. These data suggest that in the progression of cervical preneoplasia to invasive carcinomas, integration of viral genomes occurs only at single or few chromosomal loci in a given cell clone. Disruption of cellular genes might support malignant transformation in rare cases; however, it is not a pathogenic prerequisite. The main function of HPV integration seems to be the stabilization of oncogene transcription.
Ziegert
Corina
C
Institute of Molecular Pathology, Department of Pathology, University of Heidelberg, Im Neuenheimer Feld 220, 69120 Heidelberg, Germany.
Wentzensen
Nicolas
N
Vinokurova
Svetlana
S
Kisseljov
Fjodor
F
Einenkel
Jens
J
Hoeckel
Michael
M
von Knebel Doeberitz
Magnus
M
D001706:Biopsy; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000473:pathology / Q000821:virology; D002472:Cell Transformation, Viral / Q000235:genetics*; D004273:DNA, Neoplasm / Q000032:analysis*; D004279:DNA, Viral / Q000302:isolation & purification*; D018450:Disease Progression; D005260:Female; D005784:Gene Amplification; D016679:Genome, Viral; D006801:Humans; D008207:Lymphatic Metastasis; D009361:Neoplasm Invasiveness; D027383:Papillomaviridae / Q000235:genetics / Q000502:physiology*; D030361:Papillomavirus Infections / Q000235:genetics* / Q000821:virology; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014412:Tumor Virus Infections / Q000235:genetics* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology / Q000821:virology; D014625:Vaginal Neoplasms / Q000235:genetics* / Q000473:pathology / Q000821:virology; D016662:Virus Integration; D002578:Uterine Cervical Dysplasia / Q000235:genetics* / Q000473:pathology / Q000821:virology
D004273:DNA, Neoplasm; D004279:DNA, Viral; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D012367:RNA, Viral