Frequent presence of incomplete HPV16 E7 ORFs in lung carcinomas: memories of viral infection.
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.


OBJECTIVE
To examine the frequency of different length HPV16 E7 segments in lung carcinomas.


METHODS
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.
Krikelis
D
D
Department of Biological Chemistry, School of Medicine, Aristotle University of Thessaloniki, PC 54124, Thessaloniki, Greece.
Tzimagiorgis
G
G
Georgiou
E
E
Destouni
C
C
Agorastos
T
T
Haitoglou
C
C
Kouidou
S
S
D002277:Carcinoma / Q000473:pathology / Q000821:virology*; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology / Q000821:virology*; D050725:Papillomavirus E7 Proteins / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology*; D057089:Pathology, Molecular / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence
D050725:Papillomavirus E7 Proteins; C059731:oncogene protein E7, Human papillomavirus type 16