Assessing The Invasive Potential Of Bladder Cancer: Development And Validation Of A New Preclinical Assay
Bolenz C, Gorzelanny C, Knauf D, Keil T, Steidler A, Halter N, Martini T, Schneider SW. J Urol. 2012 Oct 10. pii: S0022-5347(12)05186-5. doi: 10.1016/j.juro.2012.10.007. [Epub ahead of print]

Source

Department of Urology, Mannheim Medical Center, University of Heidelberg, Mannheim, Germany. Electronic address: christian.bolenz@umm.de.

Abstract

PURPOSE:

To develop and to validate an electrophysiological method for standardized preclinical assessment of the invasive potential of urothelial carcinoma of the urinary bladder (UCB).

MATERIALS AND METHODS:

Human UCB cells (UMUC-3, RT112, HT1197 and T24/83) and benign urothelial cells (UROtsa) were cocultivated with high-resistance MDCK-C7 cells seeded below a fine pored (0.4 μm) membrane of an insert to avoid physical contact and cellular migration. Trans-epithelial electrical resistance (TEER; Ωcm2) across the MDCK-C7 monolayer was measured longitudinally and coefficients of invasive potential were calculated based on secretion of proteolytic factors by the invading cells.

RESULTS:

Consistent TEER breakdown patterns were reproduced in =14 independent samples of each cell line. Coefficients of invasive potential were significantly higher in UCB than in UROtsa cells [1.5 ± 0.32 vs. 9.9 ± 4.97 (UMUC); 12.5 ± 6.61 (T24); 20.5 ± 4.24 (RT); 21.0 ± 5.15 (HT); p<0.001]. No correlation was found between secretion patterns of matrix metalloproteinases (MMPs) -1, -2 and 9 and the invasive potential. Stimulation of UROtsa cells with recombinant human epidermal growth factor (EGF) upregulated MMP-9 secretion and significantly increased the invasive potential [1.3 ± 0.22 vs. 14.6 ± 3.28 after EGF stimulation (10 ng/ml); p<0.001].

CONCLUSIONS:

We developed a highly sensitive translational tool for studies on the initial process of metastatic spread of UCB. The presented electrophysiological invasion assay enables reliable quantification of the invasive potential of UCB cells before physical transmigration. It can be used to identify key player molecules for UCB invasion and to develop new therapeutic strategies.