[Research on 5-Fluorouracil-Induced Injury in Human Intestinal Organoids]
Jia-Rui Fang1,2, Xiao-Tong Li2, Heng Zhu2
1Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011, China.
Objective:
To investigate the damaging effect of 5-fluorouracil (5-FU) on human intestinal organoids (HIOs) derived from human induced pluripotent stem cells (hiPSCs).
Methods:
Maturely cultured HIOs were divided into a control group and a drug-induced injury group. The control group was cultured in normal medium, while the drug-induced injury group was treated with 5-FU at concentrations of 1.0, 2.0, and 4.0 μg/ml for 12, 24, and 36 hours, respectively. Immunofluorescence staining was used to detect the expression of markers (FOXA2, CDX2, KLF5) in organoids at various culture stages and to assess the expression level of the inflammatory factor IL-1β following injury. Reverse transcription-quantitative polymerase chain reaction (qRT-PCR) was used to analyze the mRNA expression changes of genes related to the organoid culture stage (FOXA2, CDX2, FOXF1) and genes related to the main intestinal cell structures (LGR5, OLFM4, MUC2, SOX9, iFABP).
Results:
Intestinal organoids derived from hiPSCs and a 5-FU-induced intestinal tissue structure injury model were successfully established. HIOs in the control group showed a good growth state and intact intestinal tissue structure; in contrast, after treatment with 1.0 μg/ml 5-FU for 36 hours, the HIOs in the drug-induced injury group exhibited significant destruction of morphological structure and a marked increase in inflammatory factor secretion.
Conclusion:
5-FU can induce structural damage to human intestinal organoids and promote the release of inflammatory factors. This study not only provides a novel experimental platform for elucidating the pathological processes of 5-FU-induced intestinal toxicity but also lays a theoretical foundation for screening protective agents and intervention strategies to mitigate chemotherapy-associated intestinal injury.
