Effect of 5-fluorouracil on mRNA expression of drug metabolizing enzyme and transporter genes in human hepatoma cell lines

Abstract

Fluoropyrimidines such as 5-fluorouracil (5-FU) are well known to have drug-drug interactions with anticoagulant medications such as warfarin. This study investigated the mRNA expression of pharmacokinetic (PK)-related genes in response to 5-FU using the hepatocarcinoma cell lines after examining relevant gene expression via RNA sequencing. We used HepaRG cells for 5-FU treatment analysis because these cells displayed PK-related gene expression. 5-FU exposure significantly reduced cytochrome P450 3A4 (CYP3A4) mRNA expression. Additionally, the mRNA expression of nuclear receptor subfamily 1 group I member 2 (also known as pregnane X receptor), a nuclear receptor transcription factor that promotes the expression of many CYP genes, was also decreased in HepaRG cells following 5-FU treatment. The mRNA expressions of the CYP2B6 and ATP-binding cassette transporter genes were decreased after 5-FU treatment. This study revealed that 5-FU treatment reduced PK-related gene expression in HepaRG cells. These findings should be useful for further drug-drug interaction research.

Changes in Pharmacodynamic Parameters during Co-administration of 5-FU with Warfarin: A Retrospective Case Series

“…[19][20][21][22][23][24][25] In our previous study using HepaRG cells to investigate 5-FU inhibition of pharmacokinetic-related gene expression, CYP3A4 mRNA levels were significantly reduced after exposure to 10 µg/mL 5-FU for 96 h. 17) CYP2C9 mRNA levels were not affected, but mRNA levels of NR1I2 (a regulator of CYPs) were also significantly reduced. 17) In contrast, the levels of CYP2C6 and CYP2C11 mRNA in rats, the functional counterparts of human CYP2C9, treated with 5-FU (120 mg/kg, single intraperitoneal injection) were significantly decreased. 28) The study used qRT-PCR using RNA extracted from rat livers 4 d after 5-FU administration.…”

“…However, the exact mechanisms of 5-FU inhibition of CYPs remain unknown. 6,7,9,11,13,17) 5-FU does not directly inhibit CYP enzymes. 18) In our previous in vitro study, HepaRG cells exposed to 5-FU at 10 µg/mL for 96 h showed significant reduction in mRNA levels of CYP3A4 and NR1I2 (a regulator of CYPs), but not in CYP2C9 mRNA.…”

“…18) In our previous in vitro study, HepaRG cells exposed to 5-FU at 10 µg/mL for 96 h showed significant reduction in mRNA levels of CYP3A4 and NR1I2 (a regulator of CYPs), but not in CYP2C9 mRNA. 17) The active metabolites of 5-FU inhibit DNA synthesis, cause DNA damage, disrupt RNA processing, and alter normal RNA function. [19][20][21][22][23][24][25] Taken together, these results suggest the possibility that 5-FU suppresses CYP protein expression by inhibiting its transcriptional regulation.…”