Quantification of β-catenin signaling components in colon cancer cell lines, tissue sections, and microdissected tumor cells using reaction monitoring mass spectrometry

Yi Chen, Mike Gruidl, Elizabeth Remily-Wood, Richard Z. Liu, Steven Eschrich, Mark Lloyd, Aejaz Nasir, Marilyn M. Bui, Emina Huang, David Shibata, Timothy Yeatman, John M. Koomen

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Reaction monitoring mass spectrometry has emerged as a powerful tool for targeted detection and quantification of proteins in clinical samples. Here, we report the use of gel electrophoresis for protein fractionation and liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM) screening for quantitative analysis of components from the Wnt/β-catenin signaling pathway, which contributes to colon tumor formation and progression. In silico tools are used to design LC-MRM screens for each target protein. Following successful peptide detection, stable isotope labeled peptides are synthesized and developed as internal standards. Then, the assays are implemented in colon cancer cell lines to achieve detection in minimal amounts of cells, compatible with direct translation to clinical specimens. Selected assays are compared with qualitative results from immunoblotting (Westerns) and translated to individual frozen colon tissue sections and laser capture microdissected tumor cells. This LC-MRM platform has been translated from in vitro models to clinical specimens, forming the basis for future experiments in patient assessment.

Original languageEnglish (US)
Pages (from-to)4215-4227
Number of pages13
JournalJournal of Proteome Research
Volume9
Issue number8
DOIs
StatePublished - Aug 6 2010

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Catenins
Colonic Neoplasms
Mass spectrometry
Tumors
Mass Spectrometry
Cells
Tissue
Cell Line
Monitoring
Assays
Colon
Neoplasms
Peptides
Wnt Signaling Pathway
Proteins
Liquid chromatography
Fractionation
Electrophoresis
Liquid Chromatography
Isotopes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

Quantification of β-catenin signaling components in colon cancer cell lines, tissue sections, and microdissected tumor cells using reaction monitoring mass spectrometry. / Chen, Yi; Gruidl, Mike; Remily-Wood, Elizabeth; Liu, Richard Z.; Eschrich, Steven; Lloyd, Mark; Nasir, Aejaz; Bui, Marilyn M.; Huang, Emina; Shibata, David; Yeatman, Timothy; Koomen, John M.

In: Journal of Proteome Research, Vol. 9, No. 8, 06.08.2010, p. 4215-4227.

Research output: Contribution to journalArticle

Chen, Y, Gruidl, M, Remily-Wood, E, Liu, RZ, Eschrich, S, Lloyd, M, Nasir, A, Bui, MM, Huang, E, Shibata, D, Yeatman, T & Koomen, JM 2010, 'Quantification of β-catenin signaling components in colon cancer cell lines, tissue sections, and microdissected tumor cells using reaction monitoring mass spectrometry', Journal of Proteome Research, vol. 9, no. 8, pp. 4215-4227. https://doi.org/10.1021/pr1005197
Chen, Yi ; Gruidl, Mike ; Remily-Wood, Elizabeth ; Liu, Richard Z. ; Eschrich, Steven ; Lloyd, Mark ; Nasir, Aejaz ; Bui, Marilyn M. ; Huang, Emina ; Shibata, David ; Yeatman, Timothy ; Koomen, John M. / Quantification of β-catenin signaling components in colon cancer cell lines, tissue sections, and microdissected tumor cells using reaction monitoring mass spectrometry. In: Journal of Proteome Research. 2010 ; Vol. 9, No. 8. pp. 4215-4227.
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