The main mechanism of 5-FU activation is conversion to fluorodeoxyuridine monophosphate (FdUMP) which inhibits the enzyme thymidylate synthase (TYMS), an important part of the folate-homocysteine cycle and purine and pyrimidine synthesis The conversion of 5-FU to FdUMP can occur via thymidylate phosphorylase (TYMP) to fluorodeoxyuridine (FUDR) and then by the action of thymidine kinase to FdUMP or indirectly via fluorouridine monophosphate (FUMP) or fluroridine (FUR) to fluorouridine diphosphate (FUDP) and then ribonucleotide reductase action to FdUDP and FdUMP. FUDP and FdUDP can also be converted to FUTP and FdUTP and incorporated into RNA and DNA respectively which also contributes to the pharmacodynamic actions of fluoropyrimidines. Sources: [https://www.pharmgkb.org/pathway/PA150653776 PharmGKB:Fluoropyrimidine Pharmacokinetics], [https://www.pharmgkb.org/pathway/PA165291507 PharmGKB:Fluoropyrimidine Pharmacodynamics], [http://en.wikipedia.org/wiki/Fluorouracil Wikipedia:Fluorouracil] Proteins on this pathway have targeted assays available via the [https://assays.cancer.gov/available_assays?wp_id=WP1601 CPTAC Assay Portal] a3c f20 a45 a42 bf9 b9d b66 b8e dd6 d42 fa0 d3c c1f ffd e0c bb0 d3c e79 cae ff5 b0a f61 c39 e25 d42 f96 aa3 dae ad5 f4c d5d a64 cf4 e3f ba0 dfc cb2 c71 d56 17695509 PubMed Human equilibrative nucleoside transporter 1, as a predictor of 5-fluorouracil resistance in human pancreatic cancer. Anticancer Res 2007 Tsujie M Nakamori S Nakahira S Takahashi Y Hayashi N Okami J Nagano H Dono K Umeshita K Sakon M Monden M 19302291 PubMed Down-regulation of cIAP2 enhances 5-FU sensitivity through the apoptotic pathway in human colon cancer cells. Cancer Sci 2009 Karasawa H Miura K Fujibuchi W Ishida K Kaneko N Kinouchi M Okabe M Ando T Murata Y Sasaki H Takami K Yamamura A Shibata C Sasaki I 15638735 PubMed Novel chemical strategies for thymidylate synthase inhibition. Curr Med Chem 2005 Gmeiner WH 18357466 PubMed Prognostic role of p53 codon 72 polymorphism in gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy. J Cancer Res Clin Oncol 2008 Huang ZH Hua D Li LH Zhu JD 15814641 PubMed Variance in the expression of 5-Fluorouracil pathway genes in colorectal cancer. Clin Cancer Res 2005 Kidd EA Yu J Li X Shannon WD Watson MA McLeod HL 18172246 PubMed Can inhibiting dihydropyrimidine dehydrogenase limit hand-foot syndrome caused by fluoropyrimidines? Clin Cancer Res 2008 Yen-Revollo JL Goldberg RM McLeod HL 19383847 PubMed Decreased levels of UMP kinase as a mechanism of fluoropyrimidine resistance. Mol Cancer Ther 2009 Humeniuk R Menon LG Mishra PJ Gorlick R Sowers R Rode W Pizzorno G Cheng YC Kemeny N Bertino JR Banerjee D 17549067 PubMed Pharmacogenetic profiling in patients with advanced colorectal cancer treated with first-line FOLFIRI chemotherapy. Pharmacogenomics J 2008 Ruzzo A Graziano F Loupakis F Santini D Catalano V Bisonni R Ficarelli R Fontana A Andreoni F Falcone A Canestrari E Tonini G Mari D Lippe P Pizzagalli F Schiavon G Alessandroni P Giustini L Maltese P Testa E Menichetti ET Magnani M 17283124 PubMed 5-Fluorouracil incorporated into DNA is excised by the Smug1 DNA glycosylase to reduce drug cytotoxicity. Cancer Res 2007 An Q Robins P Lindahl T Barnes DE 16267625 PubMed Thymidylate synthase pharmacogenetics. Invest New Drugs 2005 Marsh S 19093176 PubMed Predicting clinical outcome of 5-fluorouracil-based chemotherapy for colon cancer patients: is the CpG island methylator phenotype the 5-fluorouracil-responsive subgroup? Int J Clin Oncol 2008 Iacopetta B Kawakami K Watanabe T 19402749 PubMed Base excision by thymine DNA glycosylase mediates DNA-directed cytotoxicity of 5-fluorouracil. PLoS Biol 2009 Kunz C Focke F Saito Y Schuermann D Lettieri T Selfridge J Schär P hepatocyte CL:0000182 Cell Type 18390668 PubMed MicroRNA 29c is down-regulated in nasopharyngeal carcinomas, up-regulating mRNAs encoding extracellular matrix proteins. Proc Natl Acad Sci U S A 2008 Sengupta S den Boon JA Chen IH Newton MA Stanhope SA Cheng YJ Chen CJ Hildesheim A Sugden B Ahlquist P 18837291 PubMed Breast cancer resistance protein expression and 5-fluorouracil resistance. Biomed Environ Sci 2008 Yuan JH Cheng JQ Jiang LY Ji WD Guo LF Liu JJ Xu XY He JS Wang XM Zhuang ZX 15548681 PubMed Mechanistic and predictive profiling of 5-Fluorouracil resistance in human cancer cells. Cancer Res 2004 Wang W Cassidy J O'Brien V Ryan KM Collie-Duguid E 16510598 PubMed Pharmacogenomic identification of novel determinants of response to chemotherapy in colon cancer. Cancer Res 2006 Boyer J Allen WL McLean EG Wilson PM McCulla A Moore S Longley DB Caldas C Johnston PG 15901346 PubMed Transport mechanism and substrate specificity of human organic anion transporter 2 (hOat2 [SLC22A7]). J Pharm Pharmacol 2005 Kobayashi Y Ohshiro N Sakai R Ohbayashi M Kohyama N Yamamoto T 18267032 PubMed ERCC2 2251A>C genetic polymorphism was highly correlated with early relapse in high-risk stage II and stage III colorectal cancer patients: a preliminary study. BMC Cancer 2008 Huang MY Fang WY Lee SC Cheng TL Wang JY Lin SR 8996164 PubMed Fluorouracil in colorectal cancer--a tale of two drugs: implications for biochemical modulation. J Clin Oncol 1997 Sobrero AF Aschele C Bertino JR cancer DOID:162 Disease 14555507 PubMed Dihydropyrimidinase deficiency and severe 5-fluorouracil toxicity. Clin Cancer Res 2003 van Kuilenburg AB Meinsma R Zonnenberg BA Zoetekouw L Baas F Matsuda K Tamaki N van Gennip AH 19339911 PubMed Association of thymidylate synthase variants with 5-fluorouracil cytotoxicity. Pharmacogenet Genomics 2009 Peters EJ Kraja AT Lin SJ Yen-Revollo JL Marsh S Province MA McLeod HL 18820913 PubMed Role of BCRP as a biomarker for predicting resistance to 5-fluorouracil in breast cancer. Cancer Chemother Pharmacol 2009 Yuan J Lv H Peng B Wang C Yu Y He Z 18607850 PubMed Prognostic significance of numeric aberrations of genes for thymidylate synthase, thymidine phosphorylase and dihydrofolate reductase in colorectal cancer. Acta Oncol 2008 Jensen SA Vainer B Witton CJ Jørgensen JT Sørensen JB 16709241 PubMed Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery. Mol Cancer 2006 De Angelis PM Svendsrud DH Kravik KL Stokke T 16785472 PubMed Genetic variations in radiation and chemotherapy drug action pathways predict clinical outcomes in esophageal cancer. J Clin Oncol 2006 Wu X Gu J Wu TT Swisher SG Liao Z Correa AM Liu J Etzel CJ Amos CI Huang M Chiang SS Milas L Hittelman WN Ajani JA 18498133 PubMed Pathway based analysis of SNPs with relevance to 5-FU therapy: relation to intratumoral mRNA expression and survival. Int J Cancer 2008 Nordgard SH Alnaes GI Hihn B Lingjaerde OC Liestøl K Tsalenko A Sørlie T Lønning PE Børresen-Dale AL Kristensen VN cancer pathway PW:0000605 Pathway Ontology 2656050 PubMed Clinical pharmacology of 5-fluorouracil. Clin Pharmacokinet 1989 Diasio RB Harris BE 19144510 PubMed C677T and A1298C MTHFR polymorphisms, a challenge for antifolate and fluoropyrimidine-based therapy personalisation. Eur J Cancer 2009 De Mattia E Toffoli G 1272473 PubMed Incorporation of 3H-dibromodulcitol and 3H-dianhydrodulcitol into ascites tumor cells. Autoradiographic study. Neoplasma 1976 12724731 PubMed 5-fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 2003 Longley DB Harkin DP Johnston PG 18075467 PubMed Genetic regulation of dihydropyrimidinase and its possible implication in altered uracil catabolism. Pharmacogenet Genomics 2007 Thomas HR Ezzeldin HH Guarcello V Mattison LK Fridley BL Diasio RB drug pathway PW:0000754 Pathway Ontology 11956089 PubMed Uridine phosphorylase (-/-) murine embryonic stem cells clarify the key role of this enzyme in the regulation of the pyrimidine salvage pathway and in the activation of fluoropyrimidines. Cancer Res 2002 Cao D Russell RL Zhang D Leffert JJ Pizzorno G 9849491 PubMed Design of a novel oral fluoropyrimidine carbamate, capecitabine, which generates 5-fluorouracil selectively in tumours by enzymes concentrated in human liver and cancer tissue. Eur J Cancer 1998 Miwa M Ura M Nishida M Sawada N Ishikawa T Mori K Shimma N Umeda I Ishitsuka H 19219653 PubMed Expression profiling of nucleotide metabolism-related genes in human breast cancer cells after treatment with 5-fluorouracil. Cancer Invest 2009 Behera RK Nayak R 19077464 PubMed ATP-binding cassette C transporters in human pancreatic carcinoma cell lines. Upregulation in 5-fluorouracil-resistant cells. Pancreatology 2009 Hagmann W Jesnowski R Faissner R Guo C Löhr JM 17716232 PubMed G>C SNP of thymidylate synthase with respect to colorectal cancer. Pharmacogenomics 2007 Gusella M Padrini R 18992248 PubMed Human equilibrative nucleoside transporter 1 levels predict response to gemcitabine in patients with pancreatic cancer. Gastroenterology 2009 Farrell JJ Elsaleh H Garcia M Lai R Ammar A Regine WF Abrams R Benson AB Macdonald J Cass CE Dicker AP Mackey JR 18035049 PubMed Folylpolyglutamate synthase and gamma-glutamyl hydrolase regulate leucovorin-enhanced 5-fluorouracil anticancer activity. Biochem Biophys Res Commun 2008 Sakamoto E Tsukioka S Oie S Kobunai T Tsujimoto H Sakamoto K Okayama Y Sugimoto Y Oka T Fukushima M