title: Biological activity of probable/possible high-risk human papillomavirus types in cervical cancer.
abstract: Judging the carcinogenicity of human papillomavirus (HPV) types rarely found in cervical cancer (CxCa) is hindered by lack of studies of their biological activity in cancer tissues. To asses transcriptional activity of HPV types, we have developed ultra-short amplimer, splice-site specific, E6*I mRNA RT-PCR assays for 12 high-risk (HR)-HPV (IARC Group 1) and eight probable/possible high-risk (pHR)-HPV types (IARC Group 2A/B carcinogens). Previously unreported E6*I splice sites of the six pHR-HPV types 26, 53, 67, 70, 73 and 82 were identified by cloning and sequencing. We analyzed 97 formalin-fixed paraffin-embedded (FFPE) Mongolian CxCa biopsies for presence of HPV DNA by two sensitive genotyping assays, for E6*I transcripts of all HR-/pHR-HPV types identified and for expression of HPV surrogate markers p16(INK4a), pRb and p53. E6*I of at least one HR-/pHR-HPV was expressed in 94 (98%) of cancer tissues including seven with pHR-HPV types 26, 66, 70 or 82 as single transcribed types. Fifty-eight of E6*I mRNA transcribing cases were analyzable by immunohistochemistry and displayed p16(INK4a) overexpression in 57 (98%), pRb downregulation in 56 (97%) and p53 downregulation in 36 (62%) tissues. The newly developed E6*I mRNA RT-PCR assays appeared to be highly sensitive method to analyze HPV transcription in FFPE materials. Our finding of viral oncogene transcription of pHR-HPV types 26, 66, 70 and 82 in cervical tumors, in the absence of any other transcriptionally active HR-type and with p16(INK4a) overexpression and pRb downregulation, may support a reassessment of the carcinogenicity classification of these pHR-HPV types.
authors.affiliation: Research Program Infection and Cancer, German Cancer Research Center, DKFZ, Heidelberg, Germany.
authors.forename: Gordana
authors.initials: G
authors.lastname: Halec
authors.forename: Markus
authors.initials: M
authors.lastname: Schmitt
authors.forename: Bolormaa
authors.initials: B
authors.lastname: Dondog
authors.forename: Enkhtuya
authors.initials: E
authors.lastname: Sharkhuu
authors.forename: Nicolas
authors.initials: N
authors.lastname: Wentzensen
authors.forename: Tarik
authors.initials: T
authors.lastname: Gheit
authors.forename: Massimo
authors.initials: M
authors.lastname: Tommasino
authors.forename: Friedrich
authors.initials: F
authors.lastname: Kommoss
authors.forename: Franz X
authors.initials: FX
authors.lastname: Bosch
authors.forename: Silvia
authors.initials: S
authors.lastname: Franceschi
authors.forename: Gary
authors.initials: G
authors.lastname: Clifford
authors.forename: Lutz
authors.initials: L
authors.lastname: Gissmann
authors.forename: Michael
authors.initials: M
authors.lastname: Pawlita
mesh_terms: D000328:Adult; D000368:Aged; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D022821:RNA Splice Sites / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D016160:Retinoblastoma Protein / Q000096:biosynthesis / Q000235:genetics; D012307:Risk Factors; D014158:Transcription, Genetic / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism / Q000821:virology*
chemical_list: C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral; D009363:Neoplasm Proteins; D022821:RNA Splice Sites; D012333:RNA, Messenger; D012367:RNA, Viral; D016160:Retinoblastoma Protein; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53