2026-04-29T15:25:29Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/141302025-04-07T06:18:15Zcom_20.500.12105_15322com_20.500.12105_2051com_20.500.12105_2052col_20.500.12105_16963col_20.500.12105_16977col_20.500.12105_19617
Repisalud
author
Solis-Fernandez, Guillermo
author
Montero-Calle, Ana Maria
author
Martínez-Useros, Javier
author
López-Janeiro, Álvaro
author
de los Ríos, Vivian
author
Sanz, Rodrigo
author
Dziaková, Jana
author
Milagrosa, Elena
author
Fernández-Aceñero, María Jesús
author
Peláez-García, Alberto
author
Casal, José Ignacio
author
Hofkens, Johan
author
Rocha, Susana
author
Barderas Manchado, Rodrigo
funder
Instituto de Salud Carlos III
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
funder
Research Foundation - Flanders
funder
Ministerio de Educación, Cultura y Deporte (España)
funder
Flemish Government
2022-04-20T09:56:12Z
2022-04-20T09:56:12Z
2022-01-27
Cells. 2022;11(3):447
10.3390/cells11030447
2073-4409
Cells
35159257
http://hdl.handle.net/20.500.12105/14130
Metastasis is the primary cause of colorectal cancer (CRC) death. The liver and lung, besides adjacent lymph nodes, are the most common sites of metastasis. Here, we aimed to study the lymph nodes, liver, and lung CRC metastasis by quantitative spatial proteomics analysis using CRC cell-based models that recapitulate these metastases. The isogenic KM12 cell system composed of the non-metastatic KM12C cells, liver metastatic KM12SM cells, and liver and lung metastatic KM12L4a cells, and the isogenic non-metastatic SW480 and lymph nodes metastatic SW620 cells, were used. Cells were fractionated to study by proteomics five subcellular fractions corresponding to cytoplasm, membrane, nucleus, chromatin-bound proteins, and cytoskeletal proteins, and the secretome. Trypsin digested extracts were labeled with TMT 11-plex and fractionated prior to proteomics analysis on a Q Exactive. We provide data on protein abundance and localization of 4710 proteins in their different subcellular fractions, depicting dysregulation of proteins in abundance and/or localization in the most common sites of CRC metastasis. After bioinformatics, alterations in abundance and localization for selected proteins from diverse subcellular localizations were validated via WB, IF, IHC, and ELISA using CRC cells, patient tissues, and plasma samples. Results supported the relevance of the proteomics results in an actual CRC scenario. It was particularly relevant that the measurement of GLG1 in plasma showed diagnostic ability of advanced stages of the disease, and that the mislocalization of MUC5AC and BAIAP2 in the nucleus and membrane, respectively, was significantly associated with poor prognosis of CRC patients. Our results demonstrate that the analysis of cell extracts dilutes protein alterations in abundance in specific localizations that might only be observed studying specific subcellular fractions, as here observed for BAIAP2, GLG1, PHYHIPL, TNFRSF10A, or CDKN2AIP, which are interesting proteins that should be further analyzed in CRC metastasis.
eng
TMT
Colorectal cancer
Mass-spectrometry
Metastasis
Quantitative Proteomics
Spatial proteomics
Spatial Proteomic Analysis of Isogenic Metastatic Colorectal Cancer Cells Reveals Key Dysregulated Proteins Associated with Lymph Node, Liver, and Lung Metastasis
research article
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URL
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SpatialProteomicAnalysisIsogenic_2022.pdf
URL
https://repisalud.isciii.es/bitstreams/4d35bd6a-188e-4361-afc3-1aa26035a44e/download
File
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text/plain
SpatialProteomicAnalysisIsogenic_2022.pdf.txt