Journal: Radiology and Oncology

Article Title: The role of focal adhesion kinase in bladder cancer: translation from in vitro to ex vivo human urothelial carcinomas

doi: 10.2478/raon-2025-0052

Figure Lengend Snippet: Schematic representation of focal adhesion kinase (FAK) inhibition. We used in vitro models of a) the primary model of differentiated normal urothelium (diff NPU), in which normal porcine urothelial (NPU) cells were maintained in culture for 4 weeks, 3 weeks of which were in medium UroM (+Ca 2+ -S FBS ), b) the bladder cancer urothelium early after resection of the bladder tumour, i.e. 2-day RT4 and 2-day T24 cells (RT4 (2 d) and T24 (2 d)), and of c) the bladder cancer urothelium late after resection of the bladder tumour, i.e. 7-day RT4 and 7-day T24 cells (RT4 (7 d) and T24 (7 d)). Cells were treated with FAK inhibitors PND-1186 (1 μM and 10 μM), PF-573228 (10 μM and 100 μM) and defactinib (10 μM and 100 μM) for 2 h per day for 3 consecutive days (3 × 2 h), after which cell viability was measured.

Article Snippet: Other in vitro models include the low-grade (LG) human non-invasive papilloma urothelial cell line RT4 for NMIBC (Ta) and the high-grade (HG) human muscle-invasive cancer urothelial cell line T24 for MIBC (T2), both from ATCC (Manassas, VA).

Techniques: Inhibition, In Vitro

Journal: Radiology and Oncology

Article Title: The role of focal adhesion kinase in bladder cancer: translation from in vitro to ex vivo human urothelial carcinomas

doi: 10.2478/raon-2025-0052

Figure Lengend Snippet: Molecular characterisation of normal urothelial cells (NPU) and bladder cancer cells (RT4 and T24) in vitro . (A) Relative expression levels of UPK1B, UPK3A, CDH1, CDH2 and PTK2 were determined using qRT-PCR in differentiated NPU cells (diff NPU), 7-day RT4 and 7-day T24 cells (RT4 (7 d) and T24 (7 d)). (B) Expression of E-cadherin, N-cadherin, FAK and p-FAK in 2-day NPU cells (NPU (2 d)), diff NPU, 2-day RT4 cells (RT4 (2 d)), RT4 (7 d), 2-day T24 cells (T24 (2 d)) and T24 (7 d), determined by western blot. (C) The relative protein expression of E-cadherin, N-cadherin, FAK and p-FAK in NPU (2 d), diff NPU, RT4 (2 d), RT4 (7 d), T24 (2 d) and T24 (7 d) normalised to the expression of β-actin. The results are presented as median with interquartile range. (D) Quantification of FAK-positive RT4 (7 d) and T24 (7 d) by flow cytometry. Data are presented as the mean ± the standard error of the mean (SEM). *P < 0.05.

Article Snippet: Other in vitro models include the low-grade (LG) human non-invasive papilloma urothelial cell line RT4 for NMIBC (Ta) and the high-grade (HG) human muscle-invasive cancer urothelial cell line T24 for MIBC (T2), both from ATCC (Manassas, VA).

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry

Journal: Radiology and Oncology

Article Title: The role of focal adhesion kinase in bladder cancer: translation from in vitro to ex vivo human urothelial carcinomas

doi: 10.2478/raon-2025-0052

Figure Lengend Snippet: Effect of focal adhesion kinase (FAK) inhibitors on viabilities of differentiated normal urothelial (NPU) cells (diff NPU), 2-day RT4 and 2-day T24 cells (RT4 (2 d) and T24 (2 d)). (A) Treatment with FAK inhibitor 1 μM PND-1186 for 2 h per day for 3 consecutive days (3 × 2 h) led to a significantly lower viability of T24 (2 d) than diff NPU. (B) After 3 × 2 h treatment with 100 μM PF-573228, the viability of diff NPU was significantly higher than the viabilities of RT4 (2 d) and T24 (2 d). (C) Treatment with 100 μM defactinib for 3 × 2 h resulted in the greatest decrease in the viabilities of RT4 (2 d) and T24 (2 d) compared to the viability of diff NPU. Data are presented as the mean ± the standard error of the mean (SEM). To differentiate the statistical analysis, we compare the viability between different in vitro models (*, **, ***) and the viability between the control and the treated sample within each cell type (#). #P < 0.05. *P < 0.05. **P < 0.005. ***P < 0.001.

Article Snippet: Other in vitro models include the low-grade (LG) human non-invasive papilloma urothelial cell line RT4 for NMIBC (Ta) and the high-grade (HG) human muscle-invasive cancer urothelial cell line T24 for MIBC (T2), both from ATCC (Manassas, VA).

Techniques: In Vitro, Control

Journal: Radiology and Oncology

Article Title: The role of focal adhesion kinase in bladder cancer: translation from in vitro to ex vivo human urothelial carcinomas

doi: 10.2478/raon-2025-0052

Figure Lengend Snippet: Effect of focal adhesion kinase (FAK) inhibitors on viabilities of differentiated normal urothelial (NPU) cells (diff NPU), 7-day RT4 and 7-day T24 cells (RT4 (7 d) and T24 (7 d)). (A) After treatment with 10 μM PND-1186 for 2 h per day for 3 consecutive days (3 × 2 h), no statistically significant differences in cell viability were observed among diff NPU, RT4 (7 d) and T24 (7 d). (B) Treatment with 100 μM PF-573228 for 3 × 2 h resulted in a significant decrease in the viabilities of both RT4 (7 d) and T24 (7 d) compared to the viability of diff NPU. (C) Treatment with 100 μM defactinib for 3 × 2 h caused the greatest difference between the viabilities of diff NPU, RT4 (7 d) and T24 (7 d). (D) Western blot of p-FAK after the 3 × 2 h treatment with 100 μM defactinib in 7-day NPU cells (NPU (7 d)), RT4 (7 d) and T24 (7 d) with GAPDH-loading control. Data are presented as the mean ± the standard error of the mean (SEM). To differentiate the statistical analysis, we compare the viability between different cell types (*, **, ***) and the viability between the control and the treated sample within each cell type (#). #P < 0.05. *P < 0.05. **P < 0.005. ***P < 0.001.

Article Snippet: Other in vitro models include the low-grade (LG) human non-invasive papilloma urothelial cell line RT4 for NMIBC (Ta) and the high-grade (HG) human muscle-invasive cancer urothelial cell line T24 for MIBC (T2), both from ATCC (Manassas, VA).

Techniques: Western Blot, Control