A Proteomic Investigation to Discover Candidate Proteins Involved in Novel Mechanisms of 5-Fluorouracil Resistance in Colorectal Cancer

• Published in: Cells (Basel, Switzerland) Vol. 13; no. 4; p. 342
• Main Authors: Ortega Duran, Mario, Shaheed, Sadr ul, Sutton, Christopher W., Shnyder, Steven D.
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.02.2024
• Subjects: 5-Fluorouracil; Amino acids; Cancer therapies; Causes of; CD44 antigen; CD63 antigen; Cell culture; Cell cycle; Cell Line, Tumor; Colorectal cancer; Colorectal carcinoma; Colorectal Neoplasms - drug therapy; Colorectal Neoplasms - metabolism; Dosage and administration; Drug development; Drug resistance; drug resistance mechanisms; Drug Resistance, Neoplasm; Drug therapy; Fluorouracil; Fluorouracil - pharmacology; Fluorouracil - therapeutic use; Genomics; Health aspects; Humans; in vitro models; Labeling; Liquid chromatography; Mass spectroscopy; Metabolism; Metabolites; Mucoproteins; Oncogene Proteins; Peptides; Proteins; Proteomics; stable isotope labelling with amino acids in cell culture (SILAC); Tumorigenesis; United Kingdom; United Kingdom > UK; 5-fluorouracil; colorectal cancer; drug resistance mechanisms; proteomics; in vitro models; stable isotope labelling with amino acids in cell culture (SILAC)
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells13040342

One of the main obstacles to therapeutic success in colorectal cancer (CRC) is the development of acquired resistance to treatment with drugs such as 5-fluorouracil (5-FU). Whilst some resistance mechanisms are well known, it is clear from the stasis in therapy success rate that much is still unknown. Here, a proteomics approach is taken towards identification of candidate proteins using 5-FU-resistant sublines of human CRC cell lines generated in house. Using a multiplexed stable isotope labelling with amino acids in cell culture (SILAC) strategy, 5-FU-resistant and equivalently passaged sensitive cell lines were compared to parent cell lines by growing in Heavy medium with 2D liquid chromatography and Orbitrap Fusion™ Tribrid™ Mass Spectrometry analysis. Among 3003 commonly quantified proteins, six (CD44, APP, NAGLU, CORO7, AGR2, PLSCR1) were found up-regulated, and six (VPS45, RBMS2, RIOK1, RAP1GDS1, POLR3D, CD55) down-regulated. A total of 11 of the 12 proteins have a known association with drug resistance mechanisms or role in CRC oncogenesis. Validation through immunodetection techniques confirmed high expression of CD44 and CD63, two known drug resistance mediators with elevated proteomics expression results. The information revealed by the sensitivity of this method warrants it as an important tool for elaborating the complexity of acquired drug resistance in CRC.