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Article

Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice

  • Authors:
    • Masato Kobayashi
    • Chisana Konno
    • Ayato Shimizu
    • Masanori Kobayashi
    • Tatsuya Hori
  • View Affiliations / Copyright

    Affiliations: Laboratory of Reproduction, Nippon Veterinary and Life Science University, Musashino, Tokyo 180‑8602, Japan
  • Article Number: 60
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    Published online on: January 30, 2026
       https://doi.org/10.3892/or.2026.9065
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Abstract

Triple‑negative breast cancer (TNBC) is a highly malignant subtype with limited effective treatment options. The present study investigated the antitumor immune effects of mifepristone, a glucocorticoid receptor antagonist, using subcutaneous, orthotopic and experimental lung metastasis mouse models transplanted with 4T1 TNBC cells. Mifepristone treatment suppressed tumor growth and metastasis, leading to improved overall survival. Flow cytometric analysis of the spleen revealed decreased polymorphonuclear myeloid‑derived suppressor cells (PMN‑MDSCs) and increased T cells in the spleen, accompanied by enhanced T‑cell activity assessed ex vivo. Similar immune alterations were observed in tumor‑infiltrating cells, indicating enhanced intratumoral T‑cell responses. These results suggested that the antitumor effects of mifepristone may be partly mediated by reducing PMN‑MDSCs and restoring antitumor immunity. In tumor‑bearing mice, plasma levels of corticosterone, the major murine glucocorticoid, were elevated. In in vitro experiments using bone marrow and splenocytes, corticosterone promoted PMN‑MDSC induction and suppressed T‑cell activity, and these effects were reversed by mifepristone. Thus, mifepristone may modulate immune cell dynamics by inhibiting systemic corticosterone. To elucidate the mechanism underlying plasma corticosterone elevation, the corticosterone‑generating capacity of 4T1 cells was analyzed by exposing them to 11‑dehydrocorticosterone (DHC). The results demonstrated that 4T1 cells possessed the ability to convert the inactive form of glucocorticoid, DHC, into its active form, corticosterone, through the enzymatic activity of 11β‑hydroxysteroid dehydrogenase type 1 (11β‑HSD1). Furthermore, treatment with carbenoxolone (a non‑selective 11β‑HSD1 inhibitor) in tumor‑bearing mice decreased plasma corticosterone levels, suppressed tumor growth and produced immune changes similar to mifepristone treatment. These findings suggested that the elevated plasma corticosterone levels in tumor‑bearing mice may be mediated by 11β‑HSD1‑dependent corticosterone production, and that this mechanism was likely induced by 4T1 cells. In conclusion, the present study indicated that 4T1 cells possess corticosterone‑generating capacity through 11β‑HSD1, promoting systemic corticosterone elevation and tumor growth. Mifepristone may restore antitumor immunity, likely by reducing PMN‑MDSCs through systemic corticosterone blockade. These insights could inform the development of novel therapeutic approaches for TNBC.
View Figures

Figure 1

Mif suppresses tumor growth and
metastasis in a 4T1 breast cancer model. (A) Tumor growth curves of
4T1-Luc-bearing mice treated with Mif in the subcutaneous tumor
growth model I. 4T1-Luc cells were subcutaneously injected into the
right flank, and tumor volume was measured until day 23. Data are
presented as the mean ± SD. *P<0.01 vs. control, n=5 mice/group.
(B) Plasma corticosterone levels were measured on day 18 via tail
vein sampling in the subcutaneous tumor growth model I. n=5
mice/group. Data are presented as the mean ± SD. (C) BLI of
orthotopically implanted 4T1-Luc tumors after 3 weeks in the
orthotopic model. Quantified luciferase activity is shown. Black
bars show the median. n=5 mice/group. (D) Kaplan-Meier survival
curves for the orthotopic model. Survival rates are expressed as
the percentage of initial animals. n=5 mice/group. (E) BLI of
metastatic 4T1-Luc tumors in the experimental metastasis model.
Quantified signal intensities are shown. Black bars show the
median. n=6 mice/group. (F) Kaplan-Meier survival curve for the
experimental metastasis model. Survival rates are expressed as the
percentage of initial animals. n=6 mice/group. BLI, bioluminescent
imaging; Mif, mifepristone; Ctrl, control.

Figure 2

Mif reduces PMN-MDSCs and enhances
T-cell responses in the spleen. (A) Representative images of
spleens from sacrificed mice. (B) Correlation between tumor and
spleen weights in Ctrl and Mif-treated mice. Pearson's R and
P-values are shown. n=12 mice/group. (C) Representative flow
cytometry plots for PMN-MDSCs (CD11b+Ly6G+)
and MO-MDSCs (CD11b+Ly6C+) among splenic
CD45+ cells. (D) Percentages of PMN-MDSCs, MO-MDSCs and
total MDSCs among CD45+ cells. (E) Absolute cell numbers
of PMN-MDSCs, MO-MDSCs and total MDSCs. (F) Representative flow
cytometry plots of CD4+ and CD8+ T cells
among splenic CD45+ cells. (G) Percentages of
CD4+ and CD8+ T cells among CD45+
cells. (H) Absolute numbers of CD4+ and CD8+
T cells. (I) Ratios of CD4+ or CD8+ T cells
to total MDSCs. (J) T-cell proliferation was determined using
carboxyfluorescein succinimidyl ester dilution in CD4+
and CD8+ T cells. (K) IFN-γ levels in culture
supernatants. (A and C-K) Data were derived from the subcutaneous
tumor immunological analysis model I. (B) Data were obtained from
mice in the subcutaneous tumor growth model I and the subcutaneous
tumor immunological analysis models I and II, for which both tumor
and spleen weights were available. Data are presented as the mean ±
SD (n=4). Mif, mifepristone; Ctrl, control; PMN-MDSC,
polymorphonuclear MDSC; MO-MDSC, monocytic MDSC; MDSC,
myeloid-derived suppressor cell; IFN-γ, interferon-γ.

Figure 3

Mif decreases PMN-MDSCs and increases
T cells in the tumor microenvironment. Data were obtained from the
subcutaneous tumor immunological analysis model I. (A)
Representative flow cytometry plots of PMN-MDSCs
(CD11b+Ly6G+) and MO-MDSCs
(CD11b+Ly6C+) among tumor-infiltrating
CD45+ cells. (B) Percentages of PMN-MDSCs, MO-MDSCs and
total MDSCs among CD45+ tumor-infiltrating cells. (C)
Flow cytometry plots of tumor-infiltrating CD4+ and
CD8+ T cells. (D) Percentages of CD4+ and
CD8+ T cells among CD45+ cells. (E) Ratios of
CD4+ or CD8+ T cells to total MDSCs. Data are
presented as the mean ± SD (n=4). Mif, mifepristone; Ctrl, control;
PMN-MDSC, polymorphonuclear MDSC; MO-MDSC, monocytic MDSC; MDSC,
myeloid-derived suppressor cell.

Figure 4

Mif counteracts
corticosterone-induced T-cell suppression and MDSC expansion. (A)
Proliferation of CD4+ and CD8+ T cells
treated with corticosterone (0, 10, 100 or 500 ng/ml) in the
absence or presence of 1 µM Mif. CFSE-labeled splenocytes were
stimulated with concanavalin A for 48 h, and proliferation was
assessed using CFSE dilution. (B) Quantification of proliferating
CD4+ and CD8+ T cells. (C) IFN-γ levels in
culture supernatants. (D) Flow cytometry plots of PMN-MDSCs
(CD11b+Ly6G+) generated from bone marrow
cells cultured with granulocyte-macrophage colony-stimulating
factor and corticosterone (0, 10, 100 or 500 ng/ml) for 3 days. (E)
Percentages of PMN-MDSCs among CD45+ cells. Data are
presented as the mean ± SD (n=3). *P<0.01 vs. 0 ng/ml. Mif,
mifepristone; CFSE, carboxyfluorescein succinimidyl ester; IFN-γ,
interferon-γ; PMN-MDSC, polymorphonuclear myeloid-derived
suppressor cell.

Figure 5

Generation of corticosterone via
11β-HSD1 activity in 4T1 cells, and effects of 11β-HSD1 inhibition
on the 4T1 breast cancer model. (A) Corticosterone production by
4T1-Luc cells cultured with 100 µM CBX followed by 100 nM DHC.
Corticosterone levels in supernatants were measured using an
enzyme-linked immunosorbent assay. Data are presented as the mean ±
SD (n=3). (B) In vivo tumor growth of 4T1-Luc cells. Mice
received intraperitoneal injections of CBX, Mif or corn oil (Ctrl)
five times per week starting on day 6 post-implantation. Data are
presented as the mean ± SD. *P<0.01 vs. Ctrl. n=4 mice/group.
(C) Plasma corticosterone levels on day 14. Data are presented as
the mean±SD. (D) Percentages of PMN-MDSCs, MO-MDSCs and total MDSCs
among CD45+ cells in the spleen. (E) Percentages of
CD4+ (left) and CD8+ (right) T cells among
CD45+ cells. (F) Ratios of CD4+ or
CD8+ T cells to total MDSCs. Data are presented as the
mean ± SD (n=4). Data were derived from the (B and C) subcutaneous
tumor growth model II, and (D-F) subcutaneous tumor immunological
analysis model II. 11β-HSD1, 11β-hydroxysteroid dehydrogenase type
1; CBX, carbenoxolone; DHC, 11-dehydrocorticosterone; Mif,
mifepristone; Ctrl, control; Non-tumor control, mice without tumor
implantation; PMN-MDSC, polymorphonuclear MDSC; MO-MDSC, monocytic
MDSC; MDSC, myeloid-derived suppressor cell.
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Copy and paste a formatted citation
Spandidos Publications style
Kobayashi M, Konno C, Shimizu A, Kobayashi M and Hori T: Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice. Oncol Rep 55: 60, 2026.
APA
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., & Hori, T. (2026). Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice. Oncology Reports, 55, 60. https://doi.org/10.3892/or.2026.9065
MLA
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., Hori, T."Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice". Oncology Reports 55.4 (2026): 60.
Chicago
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., Hori, T."Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice". Oncology Reports 55, no. 4 (2026): 60. https://doi.org/10.3892/or.2026.9065
Copy and paste a formatted citation
x
Spandidos Publications style
Kobayashi M, Konno C, Shimizu A, Kobayashi M and Hori T: Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice. Oncol Rep 55: 60, 2026.
APA
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., & Hori, T. (2026). Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice. Oncology Reports, 55, 60. https://doi.org/10.3892/or.2026.9065
MLA
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., Hori, T."Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice". Oncology Reports 55.4 (2026): 60.
Chicago
Kobayashi, M., Konno, C., Shimizu, A., Kobayashi, M., Hori, T."Tumor‑associated glucocorticoid inhibition by mifepristone reduces polymorphonuclear myeloid‑derived suppressor cells and promotes antitumor immunity in triple‑negative breast cancer‑bearing mice". Oncology Reports 55, no. 4 (2026): 60. https://doi.org/10.3892/or.2026.9065
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