p53 immunodetection of liquid-based processed urinary samples helps to identify bladder tumours with a higher risk of progression

Abstract

p53 could help identify bladder tumour cases with a risk of progression from superficial to invasive disease. Semiautomatic, liquidbased cytology (LBC) techniques offer an opportunity to standardise molecular techniques. The aim of our study was to investigate whether LBC could improve p53 immunolabelling, and to assess whether urinary p53 could have a prognostic value. Immunoreactivity for p53 was studied in 198 urine samples after treatment with the Cytyc Thinprep s processor. After antigen retrieval, cells were labelled with a monoclonal antibody that recognises both wild-type and mutant form of the p53 protein (Clone DO-7, Dako), 1/1000. Positivity for p53 was assessed in 17.2% of the cases. High-grade (G3) tumours were positive in 74.1% of the cases. Comparatively, low-grade (G1 -2) urothelial carcinomas were positive in 23.5% of the cases. During a median follow-up period of 26 months, recurrence was observed in 52.9% of the cases with p53 overexpression, and in only 10.9% of negative cases (Po0.001). The progression rate was 35.3% of p53-positive cases vs 5.5% of p53-negative cases (Po0.001). Progression-free survival was significantly shorter in patients with p53 accumulation (P ¼ 0.007). In a multivariate analysis stratified on grade and stage, p53 was an independent predictor of overall survival (P ¼ 0.042). The results show that using Thinprep s LBC, p53 immunolabelling of voided urothelial cells allows most high-grade tumours to be detected and may help identify cases with a higher risk of recurrence and progression.

p16INK4a/Ki‐67 dual labelling as a marker for the presence of high‐grade cancer cells or disease progression in urinary cytopathology

“…Urinary cytology is less efficient after conservative treatment for bladder cancer, most often near 80% than 100%, and this has stimulated for years the search for biomarkers that could improve accuracy in the follow‐up period. Among many potential markers only few (including Ki‐67, uCyt+, p53 protein and FISH) have been recognized as having an impact in urinary cytology specimens 30,36,37 …”

p16INK4a overexpression and p16/Ki‐67 dual labeling versus conventional urinary cytology in the evaluation of urothelial carcinoma

“…[30][31][32][33] The sensitivity value obtained in the current series (80.8%) is not very different from those we obtained in previously published series: 83.3% and 64.1%, respectively, in new patients and in those followed after conservative treatment 31 or a global sensitivity of 88.9% in highgrade tumors. 34 The majority of series have achieved pooled sensitivity and specificity results of 42% and 96%, respectively. 35 For comparison, fluorescence in situ hybridization (FISH) using the UroVysion assay allows 72% and 83% pooled sensitivity and specificity results to be achieved.…”

“…The finding that urinary cytology is less efficient after conservative treatment for bladder cancer (most often closer to 80% than 100%) has for years stimulated the search for biomarkers that could improve accuracy in the follow‐up period. Among many potential markers, only a few (including Ki‐67, uCyt+, p53 protein, and UroVysion FISH) have been recognized as having an impact in urinary cytology specimens . In recent years, one of the most promising biomarkers has been p16 INK4a , with particular applicability in gynecopathology in the triage of ASC‐US and LSIL Pap cytology results .…”

“…Among many potential markers, only a few (including Ki-67, uCyt1, p53 protein, and UroVysion FISH) have been recognized as having an impact in urinary cytology specimens. 31,34,38 In recent years, one of the most promising biomarkers has been p16 INK4a , with particular applicability in gynecopathology in the triage of ASC-US and LSIL Pap cytology results. 19,20,39 It is also an interesting marker in other malignancies, because although CDKN2a is clearly a tumor suppressor gene, aberrant expression of its protein is observed in several human cancers.…”

p16INK4a expression in cytology of ascites and response to chemotherapy in advanced ovarian cancer

“…Because clinical trial of neoadjuvant chemotherapy in prospective setting showed positive cytology of ascites combined with image scan and serum tumor marker could confirm the diagnosis of advanced epithelial ovarian cancer for potential indication of neoadjuvant chemotherapy,5 use of ascites samples to predict chemotherpeutic response, which can be obtained in less invasive manner, seems to be promising method in those settings. In this study, we investigated p16INK4a expression as a novel prognostic marker for advanced ovarian cancer by immunocytochemistry of ascites using liquid‐based cytology samples providing complementary molecular techniques16, 17 and explored the possibility to predict the chemotherapeutic response and prognosis.…”