Detection of integrated papillomavirus sequences by ligation-mediated PCR (DIPS-PCR) and molecular characterization in cervical cancer cells

Tools

Item Type:	Article
Title:	Detection of integrated papillomavirus sequences by ligation-mediated PCR (DIPS-PCR) and molecular characterization in cervical cancer cells
Creators Name:	Luft, F.C., Klaes, R., Nees, M., Duerst, M., Heilmann, V., Melsheimer, P. and von Knebel-Doeberitz, M.
Abstract:	Human papillomavirus (HPV) genomes usually persist as episomal molecules in HPV associated preneoplastic lesions whereas they are frequently integrated into the host cell genome in HPV-related cancers cells. This suggests that malignant conversion of HPV-infected epithelia is linked to recombination of cellular and viral sequences. Due to technical limitations, precise sequence information on viral–cellular junctions were obtained only for few cell lines and primary lesions. In order to facilitate the molecular analysis of genomic HPV integration, we established a ligation-mediated PCR assay for the detectionofintegratedpapillomavirussequences (DIPS-PCR). DIPS-PCR was initially used to amplify genomic viral–cellular junctions from HPV-associated cervical cancer cell lines (C4-I, C4-II, SW756, and HeLa) and HPV-immortalized keratinocyte lines (HPKIA, HPKII). In addition to junctions already reported in public data bases, various new fusion fragments were identified. Subsequently, 22 different viral–cellular junctions were amplified from 17 cervical carcinomas and 1 vulval intraepithelial neoplasia (VIN III). Sequence analysis of each junction revealed that the viral E1 open reading frame (ORF) was fused to cellular sequences in 20 of 22 (91%) cases. Chromosomal integration loci mapped to chromosomes 1 (2n), 2 (3n), 7 (2n), 8 (3n), 10 (1n), 14 (5n), 16 (1n), 17 (2n), and mitochondrial DNA (1n), suggesting random distribution of chromosomal integration sites. Precise sequence information obtained by DIPS-PCR was further used to monitor the monoclonal origin of 4 cervical cancers, 1 case of recurrent premalignant lesions and 1 lymph node metastasis. Therefore, DIPS-PCR might allow efficient therapy control and prediction of relapse in patients with HPV-associated anogenital cancers.
Keywords:	Cervical Cancer, Human Papillomavirus, Viral Integration, Ligation-Mediated PCR, Molecular Marker
Source:	International Journal of Cancer
ISSN:	0020-7136
Publisher:	Wiley
Volume:	92
Number:	1
Page Range:	9-17
Date:	1 April 2001
Official Publication:	https://doi.org/10.1002/1097-0215(200102)9999:9999<::AID-IJC1144>3.0.CO;2-L
PubMed:	View item in PubMed

Repository Staff Only: item control page