TY - JOUR
T1 - Lipopolysaccharide-regulated secretion of soluble and vesicle-based proteins from a panel of colorectal cancer cell lines
AU - Garcia de Durango, Cira R.
AU - Monteiro, Madalena N.
AU - Bijnsdorp, Irene V.
AU - Pham, Thang V.
AU - de Wit, Meike
AU - Piersma, Sander Rogier
AU - Knol, Jaco C.
AU - Pérez-Gordo, Marina
AU - Fijneman, Remond J. A.
AU - Vidal-Vanaclocha, Fernando
AU - Jimenez, Connie R.
N1 - Funding Information:
This manuscript was supported by funding from Fundación Universitaria CEU San Pablo and Banco Santander. Cancer Center Amsterdam is acknowledged for support of the proteomics infrastructure.
Funding Information:
This manuscript was supported by funding from Fundaci?n Universitaria CEU San Pablo and Banco Santander. Cancer Center Amsterdam is acknowledged for support of the proteomics infrastructure.
Publisher Copyright:
© 2021 Wiley-VCH GmbH
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/5/1
Y1 - 2021/5/1
N2 - Purpose: To mimic the perioperative microenvironment where bacterial products get in contact with colorectal cancer (CRC) cells and study its impact on protein release, we exposed six CRC cell lines to lipopolysaccharide (LPS) and investigated the effect on the secretome using in-depth mass spectrometry-based proteomics. Experimental design: Cancer cell secretome was harvested in bio-duplicate after LPS treatment, and separated in EV and soluble secretome (SS) fractions. Gel-fractionated proteins were analysed by label-free nano-liquid chromatography coupled to tandem mass spectrometry. NF-κB activation, triggered upon LPS treatment, was evaluated. Results: We report a CRC secretome dataset of 5601 proteins. Comparison of all LPS-treated cells with controls revealed 37 proteins with altered abundance in the SS, including RPS25; and 13 in EVs, including HMGB1. Comparing controls and LPS-treated samples per cell line, revealed 564 significant differential proteins with fold-change >3. The LPS-induced release of RPS25 was validated by western blot. Conclusions and clinical relevance: Bacterial endotoxin has minor impact on the global CRC cell line secretome, yet it may alter protein release in a cell line-specific manner. This modulation might play a role in orchestrating the development of a permissive environment for CRC liver metastasis, especially through EV-communication.
AB - Purpose: To mimic the perioperative microenvironment where bacterial products get in contact with colorectal cancer (CRC) cells and study its impact on protein release, we exposed six CRC cell lines to lipopolysaccharide (LPS) and investigated the effect on the secretome using in-depth mass spectrometry-based proteomics. Experimental design: Cancer cell secretome was harvested in bio-duplicate after LPS treatment, and separated in EV and soluble secretome (SS) fractions. Gel-fractionated proteins were analysed by label-free nano-liquid chromatography coupled to tandem mass spectrometry. NF-κB activation, triggered upon LPS treatment, was evaluated. Results: We report a CRC secretome dataset of 5601 proteins. Comparison of all LPS-treated cells with controls revealed 37 proteins with altered abundance in the SS, including RPS25; and 13 in EVs, including HMGB1. Comparing controls and LPS-treated samples per cell line, revealed 564 significant differential proteins with fold-change >3. The LPS-induced release of RPS25 was validated by western blot. Conclusions and clinical relevance: Bacterial endotoxin has minor impact on the global CRC cell line secretome, yet it may alter protein release in a cell line-specific manner. This modulation might play a role in orchestrating the development of a permissive environment for CRC liver metastasis, especially through EV-communication.
KW - NF-κB
KW - colorectal cancer
KW - exosome
KW - extracellular vesicles
KW - lipopolysaccharide
KW - secretome
UR - http://www.scopus.com/inward/record.url?scp=85108303178&partnerID=8YFLogxK
U2 - 10.1002/prca.201900119
DO - 10.1002/prca.201900119
M3 - Article
C2 - 33587312
VL - 15
JO - Proteomics. Clinical applications
JF - Proteomics. Clinical applications
SN - 1862-8346
IS - 2-3
M1 - 1900119
ER -