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bioRxiv2015

Evaluation of p53 Immunohistochemical Expression Using Open-Source Software for Digital Image Analysis: A Tissue Microarray Study of Penile Squamous Cell Carcinomas

The addition of molecular biomarkers is needed to increase the accuracy of pathologic factors as prognosticators of outcome in penile squamous cell carcinomas (SCC). Evaluation of these biomarkers is usually carried out by immunohistochemistry. Herein we assess p53 immunohistochemical expression on tissue samples of penile SCC using freely-available, open-source software packages for digital image analysis. We also compared the results of digital analysis with standard visual estimation. Percentages of p53 positive cells were higher by visual estimation than by digital analysis. However, correlation was high between both methods. Our study shows that evaluation of p53 immunohistochemical expression is feasible using open-source software packages for digital image analysis. Although our analysis was limited to penile SCC, the rationale should also hold for other tumor types in which evaluation of p53 immunohistochemical expression is required. This approach would reduce interobserver variability, and would provide a standardized method for reporting the results of immunohistochemical stains. As these diagnostic tools are freely-available online, researchers and practicing pathologists could incorporate them in their daily practice without increasing diagnostic costs.

Pathology↗

bioRxiv2015

PIK3CA Mutational Analysis in Formalin-Fixed, Paraffin-Embedded Archival Tissues of Urothelial Carcinoma of Urinary Bladder

ObjectiveUrothelial carcinoma of the urinary bladder is the fourth most common cancer in males in the United States. In addition to mutations in FGFR3, TP53, AKT1, TSC1, and PTEN genes, mutations in PIK3CA have been also described in urothelial carcinomas, preferentially in low-grade tumors. Mutations in PIK3CA also has been shown to have implications for prognosis, surveillance and therapeutic response. Thus, determining the PIK3CA status in urothelial carcinomas could potentially improved the clinical management of patients with bladder cancer. Herein, we evaluated the presence of PIK3CA mutations in exons 1, 9, and 20 in 21 urothelial carcinomas of the urinary bladder.\n\nMethodsPatients were treated by radical cystectomy without neoadjuvant chemotherapy. Representative tissue blocks (1 for each case) were selected. We used a pinpoint DNA extraction technique from formalin-fixed, paraffin-embedded and mutational analysis using the polymerase chain reaction (PCR) assay coupled with sequencing of targeted exons. Patients included 15 men and 6 women, with a median age of 68 years (range, 42 to 76 years), with 3 noninvasive and 18 invasive urothelial carcinomas. Noninvasive carcinomas included 1 case each of low-grade papillary urothelial carcinoma, high-grade papillary urothelial carcinoma, and urothelial carcinoma in situ (CIS). Invasive tumors included 3 pT1, 5 pT2, 6 pT3, and 4 pT4 urothelial carcinomas.\n\nResultsWe did not find mutations in the analyzed exons of the PIK3CA gene, in any of the 21 urothelial carcinomas. The preponderance of invasive high-grade and high-stage tumors could explain the absence of identifiable mutations in our cohort.\n\nConclusionsPIK3CA mutations as prognosti-cators of outcome or predictors of therapeutic response await further evaluation.

Pathology↗