Application of artificial neural network to investigate the effects of 5-fluorouracil on ribonucleotides and deoxyribonucleotides in HepG2 cells

Abstract

Endogenous ribonucleotides and deoxyribonucleotides are essential metabolites that play important roles in a broad range of key cellular functions. Their intracellular levels could also reflect the action of nucleoside analogues. We investigated the effects of 5-fluorouracil (5-FU) on ribonucleotide and deoxyribonucleotide pool sizes in cells upon exposure to 5-FU for different durations. Unsupervised and supervised artificial neural networks were compared for comprehensive analysis of global responses to 5-FU. As expected, deoxyuridine monophosphate (dUMP) increased after 5-FU incubation due to the inhibition of thymine monophosphate (TMP) synthesis. Interestingly, the accumulation of dUMP could not lead to increased levels of deoxyuridine triphosphate (dUTP) and deoxyuridine diphosphate (dUDP). After the initial fall in intracellular deoxythymidine triphosphate (TTP) concentration, its level recovered and increased from 48 h exposure to 5-FU, although deoxythymidine diphosphate (TDP) and TMP continued to decrease compared with the control group. These findings suggest 5-FU treatment caused unexpected changes in intracellular purine polls, such as increases in deoxyadenosine triphosphate (dATP), adenosine-triphosphate (ATP), guanosine triphosphate (GTP) pools. Further elucidation of the mechanism of action of 5-FU in causing these changes should enhance development of strategies that will increase the anticancer activity of 5-FU while decreasing its resistance.

Inhibition of Orai1‐mediated Ca2+ entry enhances chemosensitivity of HepG2 hepatocarcinoma cells to 5‐fluorouracil

“…In consistent with previous studies , 5‐FU treatment for 48 hrs decreased HepG2 cell viability in a concentration‐dependent manner (Fig. A).…”

Effect of Phyllanthus amarus Extract on 5-Fluorouracil-Induced Perturbations in Ribonucleotide and Deoxyribonucleotide Pools in HepG2 Cell Line

“…When HepG2 cells were exposed to the PHA extract alone, there were no remarkably changes in the levels of the RNs, except for the slight decrease in ATP, GTP, guanosine monophosphate (GMP) and the little increase in cytidine diphosphate (CDP). The effects of 5-FU on RNs and dRNs pool sizes in cells upon exposure to 5-FU for different durations have already been reported in our previous paper [24]. The levels of four nucleoside triphosphates exhibited similar increases after treatment of 5-FU with or without PHA extract (Figure 3 and Table 2).…”

“…The ion spray voltage was set at 3000 V for negative mode and 4000 V for positive mode at a temperature of 350 °C and auxiliary gas pressure of 15 psi. Quantification was performed using multiple reactions monitoring (MRM) as previously published [24].…”

Enhanced antineoplastic/therapeutic efficacy using 5-fluorouracil-loaded calcium phosphate nanoparticles

“…An increase of 4.63-times in the apoptotic-induced cell population corroborates the previous results of the ROS induction assay, AO/EB observations, and SEM micrographs. The results infer the detrimental effect of 5-FU-induced toxicity on cell growth and multiplication [ 50 ]. The change in the mitochondrial membrane potential over the course of a 24 h treatment with CaP@5-FU NPs in HCT-15 cells was validated (Figure S5, Supporting Information File 1 ) by using JC-9 dye as the mitochondrial specific dye.…”