Prevalence of HPV genotypes in cervical adenocarcinoma: a study in Greek women.

PURPOSE: To study the prevalence of human papillomavirus (HPV) genotypes among cervical adenocarcinomas in Greek women. METHODS: The study group comprised 78 adenocarcinoma cases (20 in situ and 58 invasive). HPV DNA was amplified using polymerase chain reaction (PCR) and HPV genotypes were identified by reverse hybridization. RESULTS: There was a high prevalence of HPV infection both for in situ (95%) or invasive (94.83%) adenocarcinomas, comprising also cancers of unusual morphology. HPV 16 was the commonest strain (N=57, 73.08%) followed by HPV 18 (N=28, 35.90%). Interestingly, 13 cases (16.67%) were also HPV 52 positive (as co-infection with HPV 16 or 18). All other strains with the exception of HPV 66 were found only as co-infections. No significant age difference was noted in terms of any HPV strain positivity. CONCLUSIONS: HPV DNA was found in the large majority of cervical adenocarcinomas. As opposed to other studies, HPV 52 was the third most commonly encountered strain after HPV 16 and HPV 18. The above findings would probably be of help in decision making concerning vaccination policy for the prevention of HPV infection in Greece.

The shotgun proteomic study of the human ThinPrep cervical smear using iTRAQ mass-tagging and 2D LC-FT-Orbitrap-MS: the detection of the human papillomavirus at the protein level.

The ThinPrep cervical smear is widely used in clinical practice for the cytological and molecular screening against abnormal cells and Human Papillomavirus (HPV) infection. Current advancements made to LC-MS proteomics include the use of stable isotope labeling for the in-depth analysis of proteins in complex clinical specimens. Such approaches have yet to be realized for ThinPrep clinical specimens. In this study, an LC-MS method based on isobaric (iTRAQ) labeling and high-resolution FT-Orbitrap mass spectrometry was used for the proteomic analysis of 23 human ThinPrep smear specimens. Tandem mass spectrometry analysis was performed with both nitrogen high collision dissociation (HCD MS/MS) and helium collision induced dissociation (CID MS/MS) peptide fragmentation modes. The analysis of three 8-plex sample sets yielded the identification of over 3200 unique proteins at FDR < 1%, of which over 2300 proteins were quantitatively profiled in at least one of the three experiments. The interindividual variability served to define the required sample size needed to identify significant protein expression differences. The degree of in-depth proteome coverage allowed the detection of 6 HPV-derived proteins including the high-risk HPV16 type in the specimens tested. The presence of the HPV strains of origin was also confirmed with PCR-hybridization molecular methods. This proof-of-principle study constitutes the first ever report on the nontargeted analysis of HPV proteins in human ThinPrep clinical specimens with high-resolution mass spectrometry. A further testament to the sensitivity and selectivity of the proposed study method was the confident detection of a significant number of phosphopeptides in these specimens.