{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T21:29:48Z","timestamp":1781731788161,"version":"3.54.5"},"reference-count":21,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T00:00:00Z","timestamp":1767312000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Operational Program Integrated Infrastructure","award":["313021BUZ3"],"award-info":[{"award-number":["313021BUZ3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Background: Bladder cancer exhibits substantial heterogeneity, and accurate discrimination between non-muscle-invasive (NMIBC) and muscle-invasive disease (MIBC), as well as between low-grade (LG) and high-grade (HG) tumors, remains essential for appropriate clinical management. Established immunohistochemical (IHC) markers, such as p53, Ki-67, and E-cadherin, could be used in a new setting, but standardized cut-off values and their combined predictive value remain unclear. This study aimed to identify optimal cut-offs for these markers and to evaluate whether biomarker combinations enhance the discrimination of tumor grade and stage. Methods: A retrospective dataset of 568 cases of bladder cancer was analyzed. For each case, the expression of p53, Ki-67, and E-cadherin was quantified, and tumors were classified as NMIBC or MIBC, and as LG or HG. ROC-based cut-off selection was performed using Youden\u2019s J criterion with 10-fold stratified cross-validation. E-cadherin was modelled using an inverted scale to reflect biological loss. Logistic regression models were used to evaluate the discriminatory performance of single markers, two-marker combinations, and a three-marker model. Cross-validated AUC values and optimal thresholds were reported. Results: Ki-67 showed the strongest single-marker performance for predicting both MIBC (AUC 0.842) and HG disease (AUC 0.813), with optimal cut-offs of \u226540% and \u226530%, respectively. p53 demonstrated moderate discrimination (AUC 0.778 for MIBC and 0.776 for HG), while E-cadherin, evaluated on an inverted scale, showed acceptable performance (AUC 0.746 for MIBC; 0.780 for HG). Combining markers yielded modest improvements, with the best performance observed for Ki-67 + E-cadherin (AUCs of 0.851 for MIBC and 0.838 for HG). Conclusions: Ki-67 is the most effective single biomarker for distinguishing invasive and HG bladder cancer, while E-cadherin provides complementary value. A two-marker panel combining Ki-67 and E-cadherin, using appropriate cut-offs, offers the highest overall performance and may serve as a practical tool for enhanced pathological stratification.<\/jats:p>","DOI":"10.3390\/jcm15010338","type":"journal-article","created":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T11:11:46Z","timestamp":1767352306000},"page":"338","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Optimizing Ki-67 and E-Cadherin Thresholds for Improved Grade and Stage Classification in Urothelial Bladder Cancer"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7889-4898","authenticated-orcid":false,"given":"Stefan","family":"Harsanyi","sequence":"first","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzana Varchulova","family":"Novakova","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucia","family":"Neuschlova","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Galbavy","sequence":"additional","affiliation":[{"name":"Institute of Forensic Medicine, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5074-9621","authenticated-orcid":false,"given":"Lubos","family":"Danisovic","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stanislav","family":"Ziaran","sequence":"additional","affiliation":[{"name":"Department of Urology, Faculty of Medicine, Comenius University in Bratislava, 826 06 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarina","family":"Bevizova","sequence":"additional","affiliation":[{"name":"Institute of Anatomy, Faculty of Medicine, Comenius University in Bratislava, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,2]]},"reference":[{"key":"ref_1","unstructured":"(2025, September 29). 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