title: Comparative label-free LC-MS/MS analysis of colorectal adenocarcinoma and metastatic cells treated with 5-fluorouracil.
abstract: A label-free mass spectrometric strategy was used to examine the effect of 5-fluorouracil (5-FU) on the primary and metastatic colon carcinoma cell lines, SW480 and SW620, with and without treatment. 5-FU is the most common chemotherapeutic treatment for colon cancer. Pooled biological replicates were analyzed by nanoLC-MS/MS and protein quantification was determined via spectral counting. Phenotypic and proteomic changes were evident and often similar in both cell lines. The SW620 cells were more resistant to 5-FU treatment, with an IC(50) 2.7-fold higher than that for SW480. In addition, both cell lines showed pronounced abundance changes in pathways relating to antioxidative stress response and cell adhesion remodeling due to 5-FU treatment. For example, the detoxification enzyme NQO1 was increased with treatment in both cell lines, while disparate members of the peroxiredoxin family, PRDX2 or PRDX5 and PRDX6, were elevated with 5-FU exposure in either SW480 or SW620, respectively. Cell adhesion-associated proteins CTNNB1 and RhoA showed decreased expression with 5-FU treatment in both cell lines. The differential quantitative response in the proteomes of these patient-matched cell lines to drug treatment underscores the subtle molecular differences separating primary and metastatic cancer cells.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Kerry M
authors.initials: KM
authors.lastname: Bauer
authors.forename: Paul A
authors.initials: PA
authors.lastname: Lambert
authors.forename: Amanda B
authors.initials: AB
authors.lastname: Hummon
mesh_terms: D000230:Adenocarcinoma / Q000737:chemistry* / Q000188:drug therapy* / Q000378:metabolism; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D000975:Antioxidants; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D015179:Colorectal Neoplasms / Q000737:chemistry* / Q000188:drug therapy* / Q000378:metabolism; D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D009362:Neoplasm Metastasis; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000964:Antimetabolites, Antineoplastic; D000975:Antioxidants; D020543:Proteome; D005472:Fluorouracil