The Role of Interleukin-1-Receptor-Antagonist in Bladder Cancer Cell Migration and Invasion.
Animals
Carcinogenesis
/ genetics
Cell Line, Tumor
Cell Movement
/ genetics
Cell Proliferation
Epithelial Cells
/ metabolism
Gene Expression Regulation, Neoplastic
Humans
Interleukin 1 Receptor Antagonist Protein
/ genetics
Interleukin-1alpha
/ genetics
Interleukin-1beta
/ genetics
Neoplasm Invasiveness
Signal Transduction
Swine
Urinary Bladder
/ metabolism
Urinary Bladder Neoplasms
/ genetics
IL1RA
bladder cancer
cytokines
interleukin 1 receptor
invasion
migration
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
30 May 2021
30 May 2021
Historique:
received:
06
05
2021
revised:
21
05
2021
accepted:
27
05
2021
entrez:
2
6
2021
pubmed:
3
6
2021
medline:
24
6
2021
Statut:
epublish
Résumé
The interleukin-1-receptor antagonist IL1RA (encoded by the Based on these observations, here we investigated the potential roles of IL1RA, IL1A, and IL1B in bladder cancer cell invasion in vitro. Cell culture, real-time impedance sensing, invasion assays (Boyden chamber, pig bladder model), qPCR, Western blot, ELISA, gene overexpression. 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. 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.
Sections du résumé
BACKGROUND
BACKGROUND
The interleukin-1-receptor antagonist IL1RA (encoded by the
OBJECTIVE
OBJECTIVE
Based on these observations, here we investigated the potential roles of IL1RA, IL1A, and IL1B in bladder cancer cell invasion in vitro.
METHODS
METHODS
Cell culture, real-time impedance sensing, invasion assays (Boyden chamber, pig bladder model), qPCR, Western blot, ELISA, gene overexpression.
RESULTS
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
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.
Identifiants
pubmed: 34070905
pii: ijms22115875
doi: 10.3390/ijms22115875
pmc: PMC8198563
pii:
doi:
Substances chimiques
IL1A protein, human
0
IL1B protein, human
0
IL1RN protein, human
0
Interleukin 1 Receptor Antagonist Protein
0
Interleukin-1alpha
0
Interleukin-1beta
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Wilhelm Sander-Stiftung
ID : 2019.038.1
Références
PLoS One. 2012;7(9):e46017
pubmed: 23049925
Int J Pharm. 2009 Feb 9;367(1-2):103-8
pubmed: 18929635
J Vis Exp. 2011 Apr 01;(50):
pubmed: 21490581
Pharmacol Rep. 2015 Aug;67(4):711-8
pubmed: 26321272
PLoS One. 2012;7(10):e46536
pubmed: 23094027
Int J Mol Sci. 2021 Mar 15;22(6):
pubmed: 33803949
Int J Cancer. 2013 May 1;132(9):2032-43
pubmed: 23023397
Oncotarget. 2016 Oct 18;7(42):68527-68545
pubmed: 27602496
Cell. 2017 Oct 19;171(3):540-556.e25
pubmed: 28988769
Exp Cell Res. 2015 Jul 1;335(1):1-11
pubmed: 25911129
Biomed Opt Express. 2016 Mar 07;7(4):1193-200
pubmed: 27446646
Adv Urol. 2014;2014:184602
pubmed: 25114677
Int J Oncol. 2015 May;46(5):1978-84
pubmed: 25738940
Cancer Lett. 2016 Oct 1;380(2):534-544
pubmed: 25449784
Eur Urol. 2017 Mar;71(3):447-461
pubmed: 27324428
Acta Pharmacol Sin. 2003 Dec;24(12):1297-300
pubmed: 14653961
Ann Rheum Dis. 2000 Nov;59 Suppl 1:i60-4
pubmed: 11053091
Proc Natl Acad Sci U S A. 1991 May 1;88(9):3681-5
pubmed: 1827201
Cytotechnology. 1991 Nov;7(3):137-49
pubmed: 1368116
J Clin Invest. 1991 Nov;88(5):1445-51
pubmed: 1834696
Urol Clin North Am. 1992 Aug;19(3):499-508
pubmed: 1636234
J Urol. 2013 May;189(5):1939-44
pubmed: 23063805
J Clin Invest. 1973 Nov;52(11):2745-56
pubmed: 4355998
J Exp Med. 1995 Aug 1;182(2):623-8
pubmed: 7629520
Urology. 2006 May;67(5):1099-104
pubmed: 16698387
Mol Cancer Ther. 2016 Aug;15(8):1943-1951
pubmed: 27196763
Biomarkers. 2009 Jun;14(4):213-8
pubmed: 19489682
Analyst. 2011 Jan 21;136(2):237-45
pubmed: 20963234
World J Urol. 2019 Jan;37(1):95-105
pubmed: 30238401
Thromb Res. 2014 May;133 Suppl 2:S149-57
pubmed: 24862136
Br J Cancer. 2005 Aug 22;93(4):493-7
pubmed: 16106254
J Endourol. 2008 Nov;22(11):2547-53
pubmed: 18928386
Annu Rev Immunol. 1998;16:27-55
pubmed: 9597123
Eur Urol. 2016 Jul;70(1):106-119
pubmed: 26996659
Sci Rep. 2016 Oct 04;6:34625
pubmed: 27698389
Scand J Gastroenterol. 1997 Jun;32(6):577-81
pubmed: 9200291
Can Urol Assoc J. 2010 Feb;4(1):56-64
pubmed: 20165581
Biomed Pharmacother. 2001 Nov;55(9-10):543-7
pubmed: 11769963
Blood. 2015 May 14;125(20):3153-63
pubmed: 25977583