Genotyping of high-risk human papillomaviruses in p16/Ki-67-positive urothelial carcinoma cells: even a worm will turn.
OBJECTIVE
Co-expression of p16INK4a protein and Ki-67 (p16/Ki-67) is noted in almost all high-grade urothelial lesions. However, the aetiological role or, conversely, the absence of causative effect of high-risk human papillomaviruses (hr-HPVs) has not been documented. The purpose of this study is to evaluate HPV DNA in p16/Ki-67-positive, high-grade urothelial tumour cells.


METHODS
Fifty-seven urine samples collected from 50 patients, including 55 histologically proven high-grade proliferations and two cases with clinical evidence of malignancy, were analysed for p16/Ki-67. Immunolabelling was performed in destained Papanicolaou-stained slides after ThinPrep(®) processing. HPV genotyping was performed by polymerase chain reaction (PCR) using a DNA microarray for 35 HPV types. Confirmation of the presence (or absence) of HPV in tissue samples was verified using a reasoned approach combining PCR and in situ hybridization (ISH) for hr-HPVs.


RESULTS
Co-expression of p16/Ki-67 was noted in 43 of 57 (75.4%) cases. In these, hr-HPVs 16, 31 and 70, and low risk HPV 84, were detected in the urine in four patients (8%). Upregulation of p16INK4a protein was confirmed on bladder biopsy or transurethral resection specimens, but PCR and ISH for hr-HPVs were both negative on the tissue sections.


CONCLUSIONS
Our results show a low prevalence of HPV infection in the urinary tract of patients with p16/Ki-67-positive urothelial malignancy. The study confirms that the deregulated cell cycle, as demonstrated by p16/Ki-67 dual labelling, is independent of the oncogenic action of hr-HPVs in high-grade urothelial proliferations.
Centre de Pathologie Est, Hôpital Femme-Mère-Enfant, Hospices Civils de Lyon, Bron, France; Université Claude Bernard Lyon 1, Lyon, France.
A-S
AS
Advenier
J-S
JS
Casalegno
Y
Y
Mekki
M
M
Decaussin-Petrucci
F
F
Mège-Lechevallier
A
A
Ruffion
E
E
Piaton
D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D002295:Carcinoma, Transitional Cell / Q000737:chemistry / Q000473:pathology / Q000821:virology; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000032:analysis*; D005260:Female; D005838:Genotype; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis*; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000150:complications / Q000473:pathology; D012306:Risk; D002583:Uterine Cervical Neoplasms / Q000737:chemistry* / Q000473:pathology / Q000821:virology*
D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D019394:Ki-67 Antigen
Ki-67; bladder carcinoma; cytology; genotyping; human papillomavirus; p16INK4a; urine; urothelial carcinoma