Cell lines

HCT116 parental cell line (ATCC CCL247) and HCT116 ContinB and ContinD resistant derivatives (all wild-type TP53 cell lines)

Preparation Method

770 μM Fluorouracil (Adrucil) (5-FU) was added to the media 24 hours after seeding. Cells were harvested by trypsinization or scraping at 0, 8, and 24 hours after addition of 5-FU to the culture medium. Control wells received no Fluorouracil (Adrucil).

Reaction Conditions

770μM for 0, 8, and 24 hours

Applications

Cells were treated with Fluorouracil (Adrucil) (5-FU) for 24 hours and followed for up to 40 days in drug-free medium. After day 7, the cell counts for ContinD increased steadily, the cell counts for ContinB and parental cultures began to increase after about 15 and 20 days, respectively.

Animal models

Male C57BL/6 mice, azoxymethane/dextran sodium sulfate (AOM/DSS) model of CRC

Preparation Method

Fluorouracil (Adrucil) (5-FU) was dissolved in sterile PBS, pH7.4 and then sterile filtered through a 0.2μm syringe filter. Fluorouracil was administered in 3 cycles; cycle 1: 40mg/kg, cycle 2 and 3: 20mg/kg via intraperitoneal injection. Fluorouracil was prepared fresh at the beginning of each cycle. Sterile filtered PBS alone was used as the vehicle control. Each cycle consisted of 5 consecutive days of injections followed by 9days of recovery. The treatment period lasted for 5weeks and mice were sacrificed 24 hr after the final injection of the third cycle.

Dosage form

Intraperitoneal injection, 40mg/kg and 20mg/kg

Applications

Fluorouracil reduces tumor burden but decreases survival in AOM/DSS mice.

Fluorouracil (Adrucil) (5-FU) is an analogue of uracil with a fluorine atom at the C-5 position in place of hydrogen^[1]. Fluorouracil is widely used in the treatment of a range of cancers, including colorectal and breast cancers, and cancers of the aerodigestive tract^[1].

The mechanism of cytotoxicity of Fluorouracil (Adrucil) has been ascribed to the misincorporation of fluoronucleotides into RNA and DNA and to the inhibition of the nucleotide synthetic enzyme thymidylate synthase (TS)^[1]. Fluorouracil (Adrucil) at the concentration of 770µM treated HCT116 parental cell line and its 5-FU-resistant derivatives, Fluorouracil (Adrucil) led to a G1(/S) arrest at 8 and 24 hours^[2]. The G1 arrest was most pronounced in ContinD cells at 24 hours, whereas the S phase arrest was most pronounced in parental HCT116 cells at 24 hours^[3].

Fluorouracil (Adrucil) beneficial effected in intestinal tumorigenesis in the Apcmin/+ mice model where a 60-80% reduction in polyps^[4]. Fluorouracil (Adrucil) reduces body weight, exacerbates symptom severity score, increases liver weight, and decreases epididymal fat mass, and decrease in survival^[5].

References:
[1]. Longley D B, Harkin D P, Johnston P G. 5-fluorouracil: mechanisms of action and clinical strategies[J]. Nature reviews cancer, 2003, 3(5): 330-338.
[2]. Takeda H, Haisa M, Naomoto Y, Kawashima R, Satomoto K, Yamatuji T, Tanaka N: Effect of 5-fluorouracil on cell cycle regulatory proteins in human colon cancer cell line. Jpn J Cancer Res. 1999, 90: 677-684.
[3]. De Angelis P M, Svendsrud D H, Kravik K L, et al. Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery[J]. Molecular cancer, 2006, 5(1): 1-25.
[4]. J.M. Tucker, C. Davis, M.E. Kitchens, et al. Response to 5-fluorouracil chemotherapy is modified by dietary folic acid deficiency in Apc(Min/+) mice. Cancer Lett., 187 (2002), pp. 153-162
[5]. Sougiannis A T, VanderVeen B N, Enos R T, et al. Impact of 5 fluorouracil chemotherapy on gut inflammation, functional parameters, and gut microbiota[J]. Brain, behavior, and immunity, 2019, 80: 44-55.