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Article Open Access

Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer

  • Authors:
    • Chuanxi Lai
    • Xiya Jia
    • Yiyi Chen
    • Kangke Chen
    • Fei Wang
    • Qiqi Zhang
    • Xiaonan Xiang
    • Zhe-Sheng Chen
    • Lingna Xu
    • Sheng Dai
  • View Affiliations / Copyright

    Affiliations: Department of Colorectal Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310016, P.R. China, Bone Marrow Transplantation Center, The First Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China, Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, NY 11439, USA
    Copyright: © Lai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 4
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    Published online on: October 23, 2025
       https://doi.org/10.3892/ijmm.2025.5675
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Abstract

Despite the established clinical efficacy of oxaliplatin in colorectal cancer (CRC), resistance to this platinum‑based agent continues to pose a significant therapeutic challenge. Increased exportin 1 (XPO1) expression in CRC has been linked to chemoresistance, while KPT‑330, a selective XPO1 inhibitor, has exhibited potential in enhancing platinum drug effectiveness in other cancer types. The present study explored the synergistic effects of KPT‑330 and oxaliplatin in oxaliplatin‑resistant CRC models. Oxaliplatin‑resistant cell lines (HCT116/L‑OHP and HCT8/L‑OHP) were developed, exhibiting elevated XPO1 expression as demonstrated by western blotting. A range of in vitro assays (Cell Counting Kit‑8 assays, ethynyldeoxyuridine assays, crystal violet staining, transmission electron microscopy and flow cytometry) and an in vivo subcutaneous xenograft model in nude mice were used to evaluate the combination therapy. Co‑treatment with KPT‑330 and oxaliplatin induced G2/M phase arrest and mitochondrial dysfunction, thereby triggering apoptosis and ferroptosis. Mechanistically, the combination therapy of KPT‑330 and oxaliplatin promoted the nuclear retention of p53, which in turn upregulated p21 and downregulated solute carrier family 7 member 11. In vivo, the combination therapy significantly enhanced tumor sensitivity to oxaliplatin. These results suggested that KPT‑330 restored oxaliplatin sensitivity in resistant CRC by facilitating p53 nuclear retention, presenting a promising approach to overcome chemoresistance through dual modulation of cell cycle arrest and ferroptosis pathways.
View Figures

Figure 1

Combination of KPT-330 and
oxaliplatin synergistically inhibits the viability of
oxaliplatin-resistant CRC cells. (A) Schematic illustrating the
process of constructing oxaliplatin-resistant HCT116/L-OHP and
HCT8/L-OHP cell lines. (B) Dose-response curves for oxaliplatin in
the four CRC cell lines, with cell viability measured using the
CCK-8 assay after 48 h of treatment. (C) Immunoblot analysis of
HCT116/L-OHP and HCT8/L-OHP cells treated with oxaliplatin for 48
h. (D) Viability of four CRC cell lines and FHC cells treated with
oxaliplatin for 48 h, measured using a CCK-8 assay. (E) Viability
of indicated cancer cell lines after silencing of XPO1 or
transfection with si-NC, followed by 48 h of oxaliplatin treatment.
(F) Immunoblot analysis of HCT116/L-OHP and HCT8/L-OHP cells after
silencing of XPO1 or transfection with si-NC. (G) Dose-response
curves for KPT-330 in the four CRC cell lines and FHC cells, with
cell viability assessed using a CCK-8 assay after 48 h of
treatment. (H) Immunoblot analysis of HCT116/L-OHP and HCT8/L-OHP
cells treated with KPT-330 for 48 h. (I) Crystal violet assay for
indicated cancer cell lines treated with different dosages of
oxaliplatin and KPT-330. (J) Dose-response curves for oxaliplatin
in the two CRC cell lines with a fixed dose of KPT-330, with cell
viability measured using a CCK-8 assay after 48 h of treatment.
CCK-8, Cell Counting Kit-8; CRC, colorectal cancer; NC, negative
control; si, small interfering RNA; XPO1, exportin 1.

Figure 2

Combination of KPT-330 and
oxaliplatin induces G2/M cell cycle arrest in
oxaliplatin-resistant colorectal cancer cells. (A) Viability of
HCT116/L-OHP and HCT8/L-OHP cells treated with oxaliplatin, KPT-330
or the combination for 48 h, analyzed using a Cell Counting Kit-8
assay. (B) EdU proliferation analysis of HCT116/L-OHP and
HCT8/L-OHP cells treated with oxaliplatin, KPT-330 or the
combination for 48 h. Scale bar, 100 μm. (C) Cell cycle
distribution detected by flow cytometry after 48 h of treatment
with oxaliplatin, KPT-330 or the combination. (D) Immunoblot
analysis of HCT116/L-OHP and HCT8/L-OHP cells treated with
oxaliplatin, KPT-330 or the combination for 48 h. (E) mRNA
expression levels of CCNB1 and CDK1 in HCT116/L-OHP and HCT8/L-OHP
cells treated with oxaliplatin, KPT-330 or the combination for 48
h. **P<0.01; ***P<0.001;
****P<0.0001. CCNB1, cyclin B1; EdU,
ethynyldeoxyuridine.

Figure 3

Combination of KPT-330 and
oxaliplatin induces apoptosis in oxaliplatin-resistant colorectal
cancer cells. (A) Apoptosis rate of HCT116/L-OHP and HCT8/L-OHP
cells treated with oxaliplatin, KPT-330 or the combination for 48
h, assessed by flow cytometry. The sum of early apoptotic cells
(annexin V+/PI-) in the lower-right quadrant
and late apoptotic cells (annexin V+/PI+) in
the upper-right quadrant indicates apoptotic cells, and was used
for quantification. (B) Viability of HCT116/L-OHP and HCT8/L-OHP
cells treated with oxaliplatin and KPT-330 or the combination with
Z-VAD-FMK for 48 h, analyzed using a Cell Counting Kit-8 assay. (C)
Apoptosis rate of HCT116/L-OHP and HCT8/L-OHP cells treated with
oxaliplatin and KPT-330 or the combination with Z-VAD-FMK for 48 h,
analyzed by flow cytometry. The sum of early apoptotic cells
(annexin V+/PI-) in the lower-right quadrant
and late apoptotic cells (annexin V+/PI+) in
the upper-right quadrant indicates apoptotic cells, and was used
for quantification. (D) Immunoblot analysis of HCT116/L-OHP and
HCT8/L-OHP cells treated with oxaliplatin, KPT-330 or the
combination for 48 h. **P<0.01,
***P<0.001, ****P<0.0001. PARP, poly
(ADP-ribose) polymerase.

Figure 4

Combination of KPT-330 and
oxaliplatin increases the levels of reactive oxygen species and
induces mitochondrial dysfunction. (A) HCT116/L-OHP and HCT8/L-OHP
cells were treated with oxaliplatin, KPT-330 or the combination for
48 h, stained with JC-1, and analyzed by flow cytometry. The bar
chart shows the relative MMP loss in the four treatment groups. (B)
HCT116/L-OHP and HCT8/L-OHP cells were treated with oxaliplatin,
KPT-330 or the combination for 48 h, stained with DCFH-DA, and
analyzed by flow cytometry. (C) HCT116/L-OHP and HCT8/L-OHP cells
were treated with oxaliplatin, KPT-330 or the combination for 48 h,
stained with MitoSOX, and analyzed by flow cytometry. (D)
Representative transmission electron microscopy images of
mitochondrial morphology in HCT116/L-OHP cells treated with
oxaliplatin, KPT-330 or the combination for 48 h. Scale bars, 2
μm (upper panel) or 1 μm (lower panel).
**P<0.01, ***P<0.001,
****P<0.0001. DCFH-DA,
2',7'-dichlorodihydrofluorescein diacetate; MMP, mitochondrial
membrane potential.

Figure 5

Combination of KPT-330 and
oxaliplatin inhibits SLC7A11 and GPX4 expression to induce
ferroptosis. (A) HCT116/L-OHP and HCT8/L-OHP cells were treated
with oxaliplatin, KPT-330 or the combination for 48 h, stained with
C11-BODIPY, and analyzed by flow cytometry. (B) MDA levels after
treatment of HCT116/L-OHP and HCT8/L-OHP cells with oxaliplatin,
KPT-330 or the combination for 48 h. (C) Viability of HCT116/L-OHP
and HCT8/L-OHP cells treated with KPT-330 and oxaliplatin or the
combination with Fer-1 for 48 h, analyzed using a Cell Counting
Kit-8 assay. (D) Immunoblot analysis of HCT116/L-OHP and HCT8/L-OHP
cells treated with oxaliplatin, KPT-330 or the combination for 48
h. (E) mRNA expression levels of SLC7A11 and GPX4 in HCT116/L-OHP
and HCT8/L-OHP cells treated with oxaliplatin, KPT-330 or the
combination for 48 h. (F) Relative GSH levels assessed after
treating HCT116/L-OHP and HCT8/L-OHP cells with oxaliplatin,
KPT-330 or the combination for 48 h. (G) Immunoblot analysis of
HCT116/L-OHP cells after knockdown of p53, and treatment with
oxaliplatin, KPT-330 or the combination for 48 h.
*P<0.05, **P<0.01,
***P<0.001, ****P<0.0001. Fer-1,
ferrostatin-1; GPX4, glutathione peroxidase 4; GSH, glutathione;
MDA, malondialdehyde; si, small interfering RNA; SLC7A11, solute
carrier family 7 member 11.

Figure 6

Combination of KPT-330 and
oxaliplatin induces p53 nuclear retention. (A) HCT116/L-OHP and
HCT8/L-OHP cells were treated with oxaliplatin, KPT-330 or the
combination for 48 h, followed by collection of cytosolic and
nuclear fractions for immunoblotting analysis. (B) HCT116/L-OHP and
HCT8/L-OHP cells were treated with oxaliplatin, KPT-330 or the
combination for 48 h, and then subjected to immunofluorescence
staining. p53 localization was detected using an anti-p53 antibody,
and nuclei were labeled with DAPI. *P<0.05;
****P<0.0001. Scale bar, 50 μm.

Figure 7

Combination of KPT-330 and
oxaliplatin effectively suppresses tumor growth in a mouse
xenograft model. (A) A subcutaneous tumor xenograft model was
established by injecting HCT116/L-OHP cells into the subcutaneous
tissue of nude mice. At 1 week after tumor establishment, four
treatment groups were formed: Vehicle control, oxaliplatin, KPT-330
and combination. (B) Body weight curves of mice during the in
vivo efficacy assessment of oxaliplatin and KPT-330 in
HCT116/L-OHP xenografts. The NS symbol indicates that the weight in
the combination therapy group was not significantly different from
those in the single-drug (oxaliplatin or KPT-330) and vehicle
control groups at the final timepoint. (C) Growth curve of tumor
volume in the in vivo efficacy assessment of oxaliplatin and
KPT-330 in HCT116/L-OHP xenografts. (D) Representative images of
tumors from all groups (n=5). (E) Tumor weights in each group
(n=5). (F) Representative staining images of Ki67, H&E and p53
in HCT116/L-OHP xenograft tumors. Scale bar, 50 μm. (G)
Quantification of Ki67 in HCT116/L-OHP xenograft tumors treated
with oxaliplatin, KPT-330 or the combination. (H) Immunoblot
analysis of HCT116/L-OHP xenograft tumors treated with oxaliplatin,
KPT-330 or the combination. *P<0.05,
**P<0.01, ***P<0.001,
****P<0.0001. NS, not significant; XPO1, exportin
1.
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Copy and paste a formatted citation
Spandidos Publications style
Lai C, Jia X, Chen Y, Chen K, Wang F, Zhang Q, Xiang X, Chen Z, Xu L, Dai S, Dai S, et al: Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer. Int J Mol Med 57: 4, 2026.
APA
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q. ... Dai, S. (2026). Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer. International Journal of Molecular Medicine, 57, 4. https://doi.org/10.3892/ijmm.2025.5675
MLA
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q., Xiang, X., Chen, Z., Xu, L., Dai, S."Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer". International Journal of Molecular Medicine 57.1 (2026): 4.
Chicago
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q., Xiang, X., Chen, Z., Xu, L., Dai, S."Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer". International Journal of Molecular Medicine 57, no. 1 (2026): 4. https://doi.org/10.3892/ijmm.2025.5675
Copy and paste a formatted citation
x
Spandidos Publications style
Lai C, Jia X, Chen Y, Chen K, Wang F, Zhang Q, Xiang X, Chen Z, Xu L, Dai S, Dai S, et al: Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer. Int J Mol Med 57: 4, 2026.
APA
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q. ... Dai, S. (2026). Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer. International Journal of Molecular Medicine, 57, 4. https://doi.org/10.3892/ijmm.2025.5675
MLA
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q., Xiang, X., Chen, Z., Xu, L., Dai, S."Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer". International Journal of Molecular Medicine 57.1 (2026): 4.
Chicago
Lai, C., Jia, X., Chen, Y., Chen, K., Wang, F., Zhang, Q., Xiang, X., Chen, Z., Xu, L., Dai, S."Exportin 1 inhibitor KPT‑330 reverses oxaliplatin resistance via p53 nuclear retention in colorectal cancer". International Journal of Molecular Medicine 57, no. 1 (2026): 4. https://doi.org/10.3892/ijmm.2025.5675
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