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Viral load, gene expression and mapping of viral integration sites in HPV16‐associated HNSCC cell lines
Author(s) -
Olthof Nadine C.,
Huebbers Christian U.,
Kolligs Jutta,
Henfling Mieke,
Ramaekers Frans C.S.,
Cornet Iris,
LentAlbrechts Josefa A.,
Stegmann Alexander P.A.,
Silling Steffi,
Wieland Ulrike,
Carey Thomas E.,
Walline Heather M.,
Gollin Susanne M.,
Hoffmann Thomas K.,
Winter Johan,
Kremer Bernd,
Klussmann Jens P.,
Speel ErnstJan M.
Publication year - 2014
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.29112
Subject(s) - viral load , biology , viral oncogene , carcinogenesis , oncogene , virology , cell culture , gene , viral protein , viral structural protein , viral replication , cancer research , viral entry , virus , genetics , cell cycle
HPV‐related HNSCC generally have a better prognosis than HPV‐negative HNSCC. However, a subgroup of HPV‐positive tumors with poor prognosis has been recognized, particularly related to smoking, EGFR overexpression and chromosomal instability. Viral integration into the host genome might contribute to carcinogenesis, as is shown for cervical carcinomas. Therefore, all HPV16‐positive HNSCC cell lines currently available have been carefully analyzed for viral and host genome parameters. The viral integration status, viral load, viral gene expression and the presence of aneusomies was evaluated in the cell lines UD‐SCC‐2, UM‐SCC‐047, UM‐SCC‐104, UPCI:SCC090, UPCI:SCC152, UPCI:SCC154 and 93VU147T. HPV integration was examined using FISH, APOT‐PCR and DIPS‐PCR. Viral load and the expression of the viral genes E2, E6 and E7 were determined via quantitative PCR. All cell lines showed integration‐specific staining patterns and signals indicating transcriptional activity using FISH. APOT‐ and DIPS‐PCR identified integration‐derived fusion products in six cell lines and only episomal products for UM‐SCC‐104. Despite the observed differences in viral load and the number of viral integration sites, this did not relate to the identified viral oncogene expression. Furthermore, cell lines exhibited EGFR expression and aneusomy (except UPCI:SCC154). In conclusion, all HPV16‐positive HNSCC cell lines showed integrated and/or episomal viral DNA that is transcriptionally active, although viral oncogene expression was independent of viral copy number and the number of viral integration sites. Because these cell lines also contain EGFR expression and aneusomy, which are parameters of poor prognosis, they should be considered suitable model systems for the development of new antiviral therapies.