TY - JOUR
T1 - Methylation status of HPV16 E2-binding sites classifies subtypes of HPV-associated oropharyngeal cancers
AU - Reuschenbach, Miriam
AU - Huebbers, Christian U.
AU - Prigge, Elena-Sophie
AU - Bermejo, Justo Lorenzo
AU - Kalteis, Martin S.
AU - Preuss, Simon F.
AU - Seuthe, Inga M. C.
AU - Kolligs, Jutta
AU - Speel, Ernst-Jan M.
AU - Olthof, Nadine
AU - Kremer, Bernd
AU - Wagner, Steffen
AU - Klussmann, Jens P.
AU - Vinokurova, Svetlana
AU - Doeberitz, Magnus von Knebel
PY - 2015/6/15
Y1 - 2015/6/15
N2 - BACKGROUND The human papillomavirus (HPV) E2 protein is a transcriptional repressor of the oncogenes E6/E7 and loss of E2 function is considered a key step in carcinogenesis. Integration of HPV into the host genome may disrupt the E2 gene. Furthermore, methylation of CpG dinucleotides in E2-binding sites (E2BSs) in the HPV upstream regulatory region may interfere with transcriptional repression of E6 and E7 by E2. The authors hypothesized that the CpG methylation status of E2BS identifies subtypes of HPV type 16 (HPV16)-associated oropharyngeal squamous cell cancers (OPSCC) in association with E2 gene integrity and viral integration. METHODSMethylation of 10 CpG dinucleotides within the upstream regulatory region, encompassing E2BSs 1, 2, 3, and 4, was quantitatively analyzed by bisulfite pyrosequencing in 57 HPV16-associated OPSCC cases. E2 status was analyzed by gene amplification and quantitative real-time reverse transcriptase-polymerase chain reaction. Viral integration was determined by integration-specific polymerase chain reaction methods. RESULTSThree subgroups with differential methylation at E2BS3 and E2BS 4 were identified: 1) complete methylation (>80%) associated with the presence of integrated HPV genomes with an intact E2 gene; 2) intermediate methylation levels (20%-80%) with predominantly episomal HPV genomes with intact E2; and 3) no methylation (
AB - BACKGROUND The human papillomavirus (HPV) E2 protein is a transcriptional repressor of the oncogenes E6/E7 and loss of E2 function is considered a key step in carcinogenesis. Integration of HPV into the host genome may disrupt the E2 gene. Furthermore, methylation of CpG dinucleotides in E2-binding sites (E2BSs) in the HPV upstream regulatory region may interfere with transcriptional repression of E6 and E7 by E2. The authors hypothesized that the CpG methylation status of E2BS identifies subtypes of HPV type 16 (HPV16)-associated oropharyngeal squamous cell cancers (OPSCC) in association with E2 gene integrity and viral integration. METHODSMethylation of 10 CpG dinucleotides within the upstream regulatory region, encompassing E2BSs 1, 2, 3, and 4, was quantitatively analyzed by bisulfite pyrosequencing in 57 HPV16-associated OPSCC cases. E2 status was analyzed by gene amplification and quantitative real-time reverse transcriptase-polymerase chain reaction. Viral integration was determined by integration-specific polymerase chain reaction methods. RESULTSThree subgroups with differential methylation at E2BS3 and E2BS 4 were identified: 1) complete methylation (>80%) associated with the presence of integrated HPV genomes with an intact E2 gene; 2) intermediate methylation levels (20%-80%) with predominantly episomal HPV genomes with intact E2; and 3) no methylation (
KW - methylation
KW - human papillomavirus (HPV)
KW - upstream regulatory region
KW - E2-binding sites
KW - oropharyngeal cancer
U2 - 10.1002/cncr.29315
DO - 10.1002/cncr.29315
M3 - Article
C2 - 25731880
SN - 0008-543X
VL - 121
SP - 1966
EP - 1976
JO - Cancer
JF - Cancer
IS - 12
ER -