The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion
Background: The interleukin-1-receptor antagonist IL1RA (encoded by the <i>IL1RN</i> gene) is a potent competitive antagonist to interleukin-1 (IL1) and thereby is mainly involved in the regulation of inflammation. Previous data indicated a role of IL1RA in muscle-invasive urothelial car...
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doaj-9ca7de06ce9a4088828a6da8bbc69e662021-06-01T01:41:28ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-01225875587510.3390/ijms22115875The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and InvasionLisa Schneider0Junnan Liu1Cheng Zhang2Anca Azoitei3Sabine Meessen4Xi Zheng5Catharina Cremer6Christian Gorzelanny7Sybille Kempe-Gonzales8Cornelia Brunner9Felix Wezel10Christian Bolenz11Cagatay Gunes12Axel John13Department of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Dermatology und Venerology, UKE, 20246 Hamburg, GermanyDepartment of Otorhinolaryngology, Head and Neck Surgery, Ulm University Hospital, 89075 Ulm, GermanyDepartment of Otorhinolaryngology, Head and Neck Surgery, Ulm University Hospital, 89075 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyDepartment of Urology, Ulm University Hospital, 89081 Ulm, GermanyBackground: The interleukin-1-receptor antagonist IL1RA (encoded by the <i>IL1RN</i> gene) is a potent competitive antagonist to interleukin-1 (IL1) and thereby is mainly involved in the regulation of inflammation. Previous data indicated a role of IL1RA in muscle-invasive urothelial carcinoma of the bladder (UCB) as well as an IL1-dependent decrease in tissue barrier function, potentially contributing to cancer cell invasion. Objective: Based on these observations, here we investigated the potential roles of IL1RA, IL1A, and IL1B in bladder cancer cell invasion in vitro. Methods: Cell culture, real-time impedance sensing, invasion assays (Boyden chamber, pig bladder model), qPCR, Western blot, ELISA, gene overexpression. Results: We observed a loss of IL1RA expression in invasive, high-grade bladder cancer cell lines T24, UMUC-3, and HT1197 while IL1RA expression was readily detectable in the immortalized UROtsa cells, the non-invasive bladder cancer cell line RT4, and in benign patient urothelium. Thus, we modified the invasive human bladder cancer cell line T24 to ectopically express IL1RA, and measured changes in cell migration/invasion using the xCELLigence Real-Time-Cell-Analysis (RTCA) system and the Boyden chamber assay. The real-time observation data showed a significant decrease of cell migration and invasion in T24 cells overexpressing IL1RA (T24-IL1RA), compared to cells harboring an empty vector (T24-EV). Concurrently, tumor cytokines, e.g., IL1B, attenuated the vascular endothelial barrier, which resulted in a reduction of the Cell Index (CI), an impedance-based dimensionless unit. This reduction could be reverted by the simultaneous incubation with IL1RA. Moreover, we used an ex vivo porcine organ culture system to evaluate cell invasion capacity and showed that T24-IL1RA cells showed significantly less invasive capacity compared to parental T24 cells or T24-EV. Conclusions: Taken together, our results indicate an inverse correlation between IL1RA expression and tumor cell invasive capacity and migration, suggesting that IL1RA plays a role in bladder carcinogenesis, while the exact mechanisms by which IL1RA influences tumor cells migration/invasion remain to be clarified in future studies. Furthermore, we confirmed that real-time impedance sensing and the porcine ex vivo organ culture methods are powerful tools to discover differences in cancer cell migration and invasion.https://www.mdpi.com/1422-0067/22/11/5875bladder cancercytokinesinterleukin 1 receptorIL1RAinvasionmigration |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lisa Schneider Junnan Liu Cheng Zhang Anca Azoitei Sabine Meessen Xi Zheng Catharina Cremer Christian Gorzelanny Sybille Kempe-Gonzales Cornelia Brunner Felix Wezel Christian Bolenz Cagatay Gunes Axel John |
spellingShingle |
Lisa Schneider Junnan Liu Cheng Zhang Anca Azoitei Sabine Meessen Xi Zheng Catharina Cremer Christian Gorzelanny Sybille Kempe-Gonzales Cornelia Brunner Felix Wezel Christian Bolenz Cagatay Gunes Axel John The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion International Journal of Molecular Sciences bladder cancer cytokines interleukin 1 receptor IL1RA invasion migration |
author_facet |
Lisa Schneider Junnan Liu Cheng Zhang Anca Azoitei Sabine Meessen Xi Zheng Catharina Cremer Christian Gorzelanny Sybille Kempe-Gonzales Cornelia Brunner Felix Wezel Christian Bolenz Cagatay Gunes Axel John |
author_sort |
Lisa Schneider |
title |
The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion |
title_short |
The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion |
title_full |
The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion |
title_fullStr |
The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion |
title_full_unstemmed |
The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion |
title_sort |
role of interleukin-1-receptor-antagonist in bladder cancer cell migration and invasion |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-05-01 |
description |
Background: The interleukin-1-receptor antagonist IL1RA (encoded by the <i>IL1RN</i> gene) is a potent competitive antagonist to interleukin-1 (IL1) and thereby is mainly involved in the regulation of inflammation. Previous data indicated a role of IL1RA in muscle-invasive urothelial carcinoma of the bladder (UCB) as well as an IL1-dependent decrease in tissue barrier function, potentially contributing to cancer cell invasion. Objective: Based on these observations, here we investigated the potential roles of IL1RA, IL1A, and IL1B in bladder cancer cell invasion in vitro. Methods: Cell culture, real-time impedance sensing, invasion assays (Boyden chamber, pig bladder model), qPCR, Western blot, ELISA, gene overexpression. Results: We observed a loss of IL1RA expression in invasive, high-grade bladder cancer cell lines T24, UMUC-3, and HT1197 while IL1RA expression was readily detectable in the immortalized UROtsa cells, the non-invasive bladder cancer cell line RT4, and in benign patient urothelium. Thus, we modified the invasive human bladder cancer cell line T24 to ectopically express IL1RA, and measured changes in cell migration/invasion using the xCELLigence Real-Time-Cell-Analysis (RTCA) system and the Boyden chamber assay. The real-time observation data showed a significant decrease of cell migration and invasion in T24 cells overexpressing IL1RA (T24-IL1RA), compared to cells harboring an empty vector (T24-EV). Concurrently, tumor cytokines, e.g., IL1B, attenuated the vascular endothelial barrier, which resulted in a reduction of the Cell Index (CI), an impedance-based dimensionless unit. This reduction could be reverted by the simultaneous incubation with IL1RA. Moreover, we used an ex vivo porcine organ culture system to evaluate cell invasion capacity and showed that T24-IL1RA cells showed significantly less invasive capacity compared to parental T24 cells or T24-EV. Conclusions: Taken together, our results indicate an inverse correlation between IL1RA expression and tumor cell invasive capacity and migration, suggesting that IL1RA plays a role in bladder carcinogenesis, while the exact mechanisms by which IL1RA influences tumor cells migration/invasion remain to be clarified in future studies. Furthermore, we confirmed that real-time impedance sensing and the porcine ex vivo organ culture methods are powerful tools to discover differences in cancer cell migration and invasion. |
topic |
bladder cancer cytokines interleukin 1 receptor IL1RA invasion migration |
url |
https://www.mdpi.com/1422-0067/22/11/5875 |
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