Investigation of Noninvasive Urine Specimens with Molecular Fluctuations for a Presence of High-Risk Human Papilloma Viruses as an Inflammatory Cofactor in the Prostate Cancer

Abstract

Background: The aim of the study is to investigate noninvasive urine specimens in suspected prostate cancer (PCa) patients by a panel of PCA3, TMPRSS2-ERG fusions and GSTP1 promoter hypermethylation. Furthermore, we tested urine specimens for the presence of high-risk Human Papilloma Viruses (HPV) as an inflammatory cofactor in the complicated origin of prostate cancer (PCa). Methods: A total of 50 patients with elevated PSA and/or PCa physiological symptoms were analyzed. RNA and DNA isolation; Reverse transcription; Real-time PCR; DNA sequencing; Bisulfite conversion of DNA; Methylationspecific PCR; Cytological preparations and staining were applied. Results: Molecular fluctuations were registered in most of the patients: neoplastic GSTP1 allele, PCA3 strongly elevated expression or hyperexpression. Only in 4 cases a positive TMPRSS2-ERG status was detected. High-risk HPV types were detected in ~ 35% of our urine specimens, obtained from patients at high risk of PCa based on their molecular profiles. Approximately 96% of detected high-risk HPVs are: 16, 33, 35, 31, distributed in the subgroup with highest oncogenic potential. The estimated frequency of high-risk HPV types in control male samples with urothelial infection is significantly lower (11%). The pathological examination on cytological slides from high-risk HPV positive urine specimens showed inflammation; variable adaptations of cellular growth and differentiation and partially viral cytopathic effect. In a proportion of patients with molecular PCa disturbed profile precancerous conditions (increased primitive cells with disturbed maturation; enlarged hyperchromatic nucleus and condensed chromatin) were found. Conclusion: Our molecular PCa findings, were confirmed on the cellular level with cytological findings of high grade alterations: coarse distributed chromatin texture with nuclear membrane irregularity and thickening; high N:C Ratio; prominence of nucleoli and irregularity in shape thereof; identical monotonous nucleoli present in all cells in a group (i.e., "Clonal" pattern); Tumor diathesis. The present study concerns novel data for Bulgarian PCa patients.