Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy

Jun LI; Zongyuan XIE

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Jun LI, Zongyuan XIE. Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy[J]. Journal of Kunming Medical University.

Citation:

Jun LI, Zongyuan XIE. Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy[J]. Journal of Kunming Medical University.

Jun LI, Zongyuan XIE. Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy[J]. Journal of Kunming Medical University.

Citation:

Jun LI, Zongyuan XIE. Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy[J]. Journal of Kunming Medical University.

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Mechanism Study of SIRT3 Enhancing Chemosensitivity of Rectal Cancer Cells to 5-Fluorouracil via FoxO3a/BNIP3-Mediated Mitophagy

Jun LI¹,
Zongyuan XIE^{2
,
,}

1.
Pharmacy Department，Linxi Hospital，Kailuan General Hospital，Tangshan 063103
2.
Magnetic Resonance Room，Affiliated Hospital of North China University of Science and Technology

Received Date: 2025-12-16
Available Online: 2026-04-28

Abstract

Abstract

Objective To investigate the role of Sirtuin 3 （SIRT3） in 5-fluorouracil （5-FU） resistance of colorectal cancer （CRC） cells and to elucidate whether it enhances tumor cell sensitivity to 5-FU by regulating Forkhead box O3a （FoxO3a）/BCL2 interacting protein 3 （BNIP3）-mediated mitophagy. Methods A 5-FU-resistant CRC cell line was established, and a SIRT3 stably overexpressing cell line was constructed via lentiviral transfection. The half-maximal inhibitory concentration （IC₅₀） was measured by CCK-8 assay, and apoptosis was detected by flow cytometry. Western blot was used to detect the expression of SIRT3, FoxO3a, BNIP3, and mitophagy-associated proteins including microtubule-associated protein 1 light chain 3 （LC3）, sequestosome 1 （p62）, PTEN-induced putative kinase 1 （PINK1）, and Parkin RBR E3 ubiquitin-protein ligase （Parkin）. Co-localization of mitochondria and autophagosomes was observed by immunofluorescence staining. Western blot was used to detect the expression of SIRT3, FoxO3a, BNIP3, and mitophagy-associated proteins including microtubule-associated protein 1 light chain 3 (LC3), sequestosome 1 (p62), PTEN-induced putative kinase 1 (PINK1), Parkin RBR E3 ubiquitin-protein ligase (Parkin) and FoxO3a acetylation level. A subcutaneous xenograft tumor model in nude mice was established to evaluate the anti-tumor effect of SIRT3 overexpression in vivo. Results The protein expression of SIRT3, FoxO3a, and BNIP3 was significantly downregulated in 5-FU-resistant CRC cells compared with parental cells （P < 0.05）, accompanied by decreased co-localization of mitochondria and autophagosomes. Overexpression of SIRT3 markedly reduced the IC₅₀ value of 5-FU-resistant cells, increased the apoptosis rate （P < 0.01）, elevated the LC3-II/I ratio, decreased p62 expression, and upregulated PINK1 and Parkin expression （P < 0.05）. Furthermore, SIRT3 overexpression decreased FoxO3a acetylation and increased BNIP3 expression. The combination of SIRT3 overexpression and 5-FU significantly inhibited tumor growth in vivo （P < 0.01）, and the changes in autophagy-associated protein expression in tumor tissues were consistent with the in vitro results, with an increase in autophagosomes. Conclusion SIRT3 reverses 5-FU resistance in CRC cells by deacetylating and activating the FoxO3a/BNIP3 pathway to induce mitophagy, and combination therapy exerts a synergistic anti-tumor effect.
- Colorectal cancer,
- SIRT3,
- 5-Fluorouracil,
- Chemoresistance,
- Mitophagy,
- FoxO3a,
- BNIP3

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References(24)

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