Assessing the Invasive Potential of Bladder Cancer: Development and Validation of a New Preclinical Assay
| Autor: | Christian Bolenz, Thomas Martini, Daniel Knauf, Tanja Keil, Annette Steidler, Stefan W. Schneider, Natalia Halter, Christian Gorzelanny |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Carcinoma
Transitional Cell medicine.medical_specialty Bladder cancer Urinary bladder business.industry Urology Cell migration medicine.disease Extracellular matrix medicine.anatomical_structure Urinary Bladder Neoplasms Cell culture Epidermal growth factor Electric Impedance Tumor Cells Cultured medicine Carcinoma Humans Biological Assay Neoplasm Invasiveness Urothelium business |
| Zdroj: | Journal of Urology. 189:1939-1944 |
| ISSN: | 1527-3792 0022-5347 |
| DOI: | 10.1016/j.juro.2012.10.007 |
| Popis: | We developed and validated an electrophysiological method for standardized preclinical assessment of the invasive potential of urothelial carcinoma of the bladder.Human UMUC-3, RT-112, HT-1197 and T24/83 bladder urothelial carcinoma cells, and UROtsa benign urothelial cells were co-cultivated with high resistance MDCK-C7 cells seeded below a 0.4 μm pore membrane of an insert to avoid physical contact and cellular migration. Transepithelial electrical resistance in Ω cm(2) across the MDCK-C7 monolayer was measured longitudinally. Invasive potential coefficients were calculated based on the secretion of proteolytic factors by invading cells.Consistent transepithelial electrical resistance breakdown patterns were reproduced in 14 or more independent samples of each cell line. Coefficients of invasive potential were significantly higher in bladder urothelial carcinoma than UROtsa cells, including a mean ± SD of 1.5 ± 0.32 vs 9.9 ± 4.97 in UMUC-3, 12.5 ± 6.61 in T24/83, 20.5 ± 4.24 in RT-112 and 21.0 ± 5.15 in HT-1197 cells (p0.001). No correlation was found between the secretion patterns of matrix metalloproteinase-1, 2 and 9, and invasive potential. Stimulation of UROtsa cells with recombinant human epidermal growth factor up-regulated matrix metalloproteinase-9 secretion and significantly increased invasive potential a mean of 1.3 ± 0.22 vs 14.6 ± 3.28 after stimulation with 10 ng/ml epidermal growth factor (p0.001).We developed a highly sensitive translational tool to study the initial process of metastatic spread of urothelial carcinoma of the bladder. The presented electrophysiological invasion assay enables reliable quantification of the invasive potential of bladder urothelial carcinoma cells before physical transmigration. It can be used to identify key molecules for bladder urothelial carcinoma invasion and develop new therapeutic strategies. |
| Databáze: | OpenAIRE |
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