BACKGROUND: HPV16 E6/E7 oncoproteins are critical for cervical carcinogenesis. The corresponding oncogenes are also detected in head and neck cancer, but in lung cancer their presence is strongly debated. PCR-based detection protocols amplify different target sequences. OBJECTIVES: To examine the frequency of different length HPV16 E7 segments in lung carcinomas. STUDY DESIGN: We designed four different amplification schemes for the detection of overlapping segments of the HPV16 E7 ORF, all suitable for specific HPV detection in cervical carcinoma. In two schemes, the entire E7 ORF was targeted while in the remaining schemes internal, smaller sequences were targeted. In total, 76 specimens were used; 29 lung carcinoma specimens, 16 non-cancerous lung tissue specimens from the same patients and 31 bronchial washings from different lung cancer patients. RESULTS: Amplification of the entire HPV16 E7 ORF, using two protocols, demonstrated the absence of the specific HPV16 E7 sequences (74 samples either tested negative by the first PCR protocol or false positive by the second, based on sequencing or AvaII or PvuII digestion). However, both schemes targeting smaller E7 segments revealed the frequent presence of HPV16 E7 sequences in lung carcinoma specimens (14/23 positive by either scheme). CONCLUSIONS: HPV16 E7 sequences are frequently observed in lung carcinomas. Decreasing the size of PCR-target sequences increases the detection frequency, possibly indicating the presence of incomplete viral ORFs. Restriction endonuclease analysis is critical for verifying the reliability of the detection of these sequences.
BACKGROUND:HPV16 E6/E7 oncoproteins are critical for cervical carcinogenesis. The corresponding oncogenes are also detected in head and neck cancer, but in lung cancer their presence is strongly debated. PCR-based detection protocols amplify different target sequences. OBJECTIVES: To examine the frequency of different length HPV16 E7 segments in lung carcinomas. STUDY DESIGN: We designed four different amplification schemes for the detection of overlapping segments of the HPV16 E7 ORF, all suitable for specific HPV detection in cervical carcinoma. In two schemes, the entire E7 ORF was targeted while in the remaining schemes internal, smaller sequences were targeted. In total, 76 specimens were used; 29 lung carcinoma specimens, 16 non-cancerous lung tissue specimens from the same patients and 31 bronchial washings from different lung cancerpatients. RESULTS: Amplification of the entire HPV16 E7 ORF, using two protocols, demonstrated the absence of the specific HPV16 E7 sequences (74 samples either tested negative by the first PCR protocol or false positive by the second, based on sequencing or AvaII or PvuII digestion). However, both schemes targeting smaller E7 segments revealed the frequent presence of HPV16 E7 sequences in lung carcinoma specimens (14/23 positive by either scheme). CONCLUSIONS:HPV16 E7 sequences are frequently observed in lung carcinomas. Decreasing the size of PCR-target sequences increases the detection frequency, possibly indicating the presence of incomplete viral ORFs. Restriction endonuclease analysis is critical for verifying the reliability of the detection of these sequences.
Authors: Camille Ragin; Monisola Obikoya-Malomo; Sungjin Kim; Zhengjia Chen; Rafael Flores-Obando; Denise Gibbs; Chihaya Koriyama; Francisco Aguayo; Jill Koshiol; Neil E Caporaso; Giovanna E Carpagnano; Marco Ciotti; Hirotoshi Dosaka-Akita; Masashi Fukayama; Akiteru Goto; Demetrios A Spandidos; Vassilis Gorgoulis; Daniëlle A M Heideman; Robert A A van Boerdonk; Kenzo Hiroshima; Reika Iwakawa; Nikolaos G Kastrinakis; Ichiro Kinoshita; Suminori Akiba; Maria T Landi; H Eugene Liu; Jinn-Li Wang; Ranee Mehra; Fadlo R Khuri; Wan-Teck Lim; Taofeek K Owonikoko; Suresh Ramalingam; Emmanuela Sarchianaki; Kari Syrjanen; Ming-Sound Tsao; Jenna Sykes; Siew Wan Hee; Jun Yokota; Apostolos Zaravinos; Emanuela Taioli Journal: Carcinogenesis Date: 2014-02-12 Impact factor: 4.944
Authors: Juan Pablo Muñoz; Carolina González; Bárbara Parra; Alejandro H Corvalán; Maria Lina Tornesello; Yoshito Eizuru; Francisco Aguayo Journal: PLoS One Date: 2012-05-25 Impact factor: 3.240