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Epidermal growth factor receptor function in the human urothelium

Artikel i vetenskaplig tidskrift
Författare Caroline Wasén
Matias Ekstrand
Max Levin
Daniel Giglio
Publicerad i International Urology and Nephrology
Volym 50
Nummer/häfte 4
Sidor 647-656
ISSN 0301-1623
Publiceringsår 2018
Publicerad vid Wallenberglaboratoriet
Institutionen för kliniska vetenskaper, Avdelningen för onkologi
Institutionen för neurovetenskap och fysiologi, sektionen för farmakologi
Sidor 647-656
Språk en
Länkar https://doi.org/10.1007/s11255-018-...
Ämnesord Epidermal growth factor receptor, Occludin, Proliferation, Three-dimensional cell culture, Urothelium
Ämneskategorier Farmakologi

Sammanfattning

© 2018, The Author(s). Purpose: Epidermal growth factor receptor (EGFr)-targeted therapy may be used in subgroups of patients with urinary bladder cancer. Here we assessed the role of EGFr in urothelial proliferation and migration in a two- and three-dimensional cell culture system. Methods: UROtsa cells derived from normal urothelium and malignant T24 cells were cultured in a Type I collagen gel. Proliferation and migration of urothelial cells, in the absence and presence of the EGFr inhibitor cetuximab, were assessed with a proliferation test (ATCC) and with the Axioplan 2 imaging microscope with a motorized stage (Carl Zeiss), respectively. The expressions of cytokeratin (CK) 17, CK20, EGFr, pEGFr, laminin, occludin and zonula occludens 1 (ZO-1) were assessed with immunohistochemistry and/or western blot. Results: UROtsa spheroids were formed after 7 days in culture, while T24 cells did not form spheroids. UROtsa expressed CK20 but not laminin or CK17 and consequently resembled umbrella cells. In UROtsa and T24, cetuximab inhibited urothelial proliferation, induced cleavage of EGFr and/or pEGFR but did not affect urothelial migration. The tight junction protein occludin was cleaved, and the formation of cellular spheroids was inhibited in UROtsa by the presence of cetuximab. Conclusions: EGFr modulates urothelial proliferation and the formation of the three-dimensional structure of the urothelium possibly by interfering with occludin. The present data also show a cell culture technique enabling phenotypically normal urothelial cells to form epithelial structures in contrast to malignant urothelial cells.

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