title: Suppression of TWIST1 enhances the sensitivity of colon cancer cells to 5-fluorouracil.
abstract: The 5-fluorouracil (5FU)-based adjuvant chemotherapy improves the survival of patients with colorectal cancer, however the main obstacle affecting its effectiveness is a drug resistance. Our study aimed to investigate the impact of TWIST1 silencing on the sensitivity of cancer cells to 5FU. The suppression of TWIST1 expression in human colon cancer HT29 and HCT116 cell lines was achieved by transduction with lentiviral vector carrying the TWIST1 silencing sequence (pLL3.7-shTWIST1). The suppression of TWIST1 expression induced changes in the expression pattern of epithelial to mesenchymal transition markers, reduced the cells proliferation rate, increased their sensitivity to serum withdrawn, and increased the cytotoxic effect of 5FU. However, significantly higher 5FU cytotoxicity was observed in HT29 cell cultures. Cells with silenced TWIST1 displayed altered expression of enzymes metabolizing 5FU. The expression level of dihydropyrimidine dehydrogenase, and thymidylate synthase decreased significantly in HT29 shTWIST1 cells, but not in HCT116 shTWIST1 cells. On the other hand, significant increases in the expression levels of thymidine phosphorylase, and uridine phosphorylase 1 were seen in both cell lines with suppressed expression of TWIST1. The changes in enzymes expression were mirrored by enzymatic activities. In conclusion, our observations point to TWIST1 as a target protein to enhance the sensitivity of colorectal cancer cells to 5FU.
authors.affiliation: Department of Molecular Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Sakowicz-Burkiewicz
authors.affiliation: Department of Molecular Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Przybyla
authors.affiliation: Department of Molecular Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland.
authors.forename: Martyna
authors.initials: M
authors.lastname: Wesserling
authors.affiliation: Departemnt of Laboratory Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland.
authors.forename: Hanna
authors.initials: H
authors.lastname: Bielarczyk
authors.affiliation: Department of Molecular Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland.
authors.forename: Izabela
authors.initials: I
authors.lastname: Maciejewska
authors.affiliation: Department of Molecular Medicine, Medical University of Gdansk, 80-211 Gdansk, Poland. Electronic address: tkpaw@gumed.edu.pl.
authors.forename: Tadeusz
authors.initials: T
authors.lastname: Pawelczyk
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D003110:Colonic Neoplasms / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D005472:Fluorouracil / Q000378:metabolism / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing; D045325:HCT116 Cells; D019073:HT29 Cells; D006801:Humans; D051879:Twist-Related Protein 1 / Q000172:deficiency* / Q000235:genetics*
chemical_list: D051879:Twist-Related Protein 1; D005472:Fluorouracil
keywords: 5-fluorouracil; Colon cancer; DPYD; TWIST1; TYMP; UPP1