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Quantitative proteomics of fractionated membrane and lumen exosome proteins from isogenic metastatic and nonmetastatic bladder cancer cells reveal differential expression of EMT factors

Jeppesen, Dennis Kjølhede, Nawrocki, Arkadiusz, Jensen, Steffen Grann, Thorsen, Kasper, Whitehead, Bradley, Howard, Kenneth A., Dyrskjøt, Lars, Ørntoft, Torben Falck, Larsen, Martin R., Ostenfeld, Marie Stampe·2014·PROTEOMICS

Abstract

Cancer cells secrete soluble factors and various extracellular vesicles, including exosomes, into their tissue microenvironment. The secretion of exosomes is speculated to facilitate local invasion and metastatic spread. Here, we used an in vivo metastasis model of human bladder carcinoma cell line T24 without metastatic capacity and its two isogenic derivate cell lines SLT4 and FL3, which form metastases in the lungs and liver of mice, respectively. Cultivation in CLAD1000 bioreactors rather than conventional culture flasks resulted in a 13‐ to 16‐fold increased exosome yield and facilitated quantitative proteomics of fractionated exosomes. Exosomes from T24, SLT4, and FL3 cells were partitioned into membrane and luminal fractions and changes in protein abundance related to the gain of metastatic capacity were identified by quantitative iTRAQ proteomics. We identified several proteins linked to epithelial–mesenchymal transition, including increased abundance of vimentin and hepatoma‐derived growth factor in the membrane, and casein kinase II α and annexin A2 in the lumen of exosomes, respectively, from metastatic cells. The change in exosome protein abundance correlated little, although significant for FL3 versus T24, with changes in cellular mRNA expression. Our proteomic approach may help identification of proteins in the membrane and lumen of exosomes potentially involved in the metastatic process.

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Metadata

Authors
Jeppesen, D. K., Nawrocki, A., Jensen, S. G., Thorsen, K., Whitehead, B., Howard, K. A., Dyrskjøt, L., Ørntoft, T. F., Larsen, M. R., & Ostenfeld, M. S.
Venue
PROTEOMICS
Publisher
Wiley
Published
2014-02-16
Volume
14
Issue
6
Pages
699-712
ISSN
  • 1615-9853 (ISSN-L)
  • 1615-9861
OpenAlex
W2071134774
Cited by
177
References
10
FWCI
5.78
Type
journal article
Language
en
Update notices
erratum (2014-11-12)
Open access
closed

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CoChat. (2026). Citation check: Quantitative proteomics of fractionated membrane and lumen exosome proteins from isogenic metastatic and nonmetastatic bladder cancer cells reveal differential expression of EMT factors. https://app.cochat.ai/papers/10.1002/pmic.201300452
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Jeppesen, D. K., Nawrocki, A., Jensen, S. G., Thorsen, K., Whitehead, B., Howard, K. A., Dyrskjøt, L., Ørntoft, T. F., Larsen, M. R., & Ostenfeld, M. S. (2014). Quantitative proteomics of fractionated membrane and lumen exosome proteins from isogenic metastatic and nonmetastatic bladder cancer cells reveal differential expression of EMT factors. PROTEOMICS. https://doi.org/10.1002/pmic.201300452

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