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Article Open Access

Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction

  • Authors:
    • Ryo Saga
    • Yoichiro Hosokawa
    • Manabu Fukumoto
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    Affiliations: Department of Radiation Science, Hirosaki University Graduate School of Health Sciences, Hirosaki, Aomori 036‑8564, Japan, Department of Rehabilitation Science, Hirosaki University of Health and Welfare, Hirosaki, Aomori 036‑8102, Japan, Radiological Disasters and Medical Science Laboratory, Disaster Medical Science Division, International Research Institute of Disaster Science, Tohoku University, Sendai, Miyagi 980‑8572, Japan
    Copyright: © Saga et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 51
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    Published online on: August 11, 2026
       https://doi.org/10.3892/mi.2026.335
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Abstract

Numerous patients with cancer experience metastasis, recurrence and resistance to chemotherapy and radiotherapy due to the presence of cancer stem cells (CSCs). Salinomycin (Sal) is a candidate for CSC‑targeting therapy and may consequently sensitize tumors to radiation. Therefore, the present study investigated whether Sal can alleviate radioresistance using a clinically relevant radioresistant cell line and sorted CSC‑like cells. The human oral squamous carcinoma cell line, SAS, and its radioresistant counterpart, SAS‑R, were used. The fraction of CD24‑/CD44+ cells was identified in the CSC‑like cell population and that of CD24+/CD44+ cells was in identified in the non‑CSC‑like counterparts. These were sorted from the SAS cells. Clonogenic potency was evaluated using a colony formation assay. The positive fractions of CSC markers (CD24 and CD44) and the number of apoptotic SAS and SAS‑R cells were measured using flow cytometry. The percentage of the CD24+/CD44+ fraction in the SAS‑R cells was lower than that in the SAS cells, whereas the CD24‑/CD44+ population was higher in the SAS‑R cells than in the SAS cells. The Sal concentration‑dependent decrease in the cell surviving fraction was more rapid in the SAS‑R cells than in the SAS cells (IC50 values: 0.250 and 0.332 µM, respectively). The fraction of CD24‑/CD44+ cells exhibited a greater radioresistance than the parent SAS cells and the fraction of CD24+/CD44+ cells. Moreover, the surviving fraction of CD24‑/CD44+ cells was significantly reduced by treatment with 0.3 µM Sal. The addition of Sal increased the number of apoptotic cells compared to 2 Gy radiation alone. On the whole, the present study indicates that Sal is effective in killing the radioresistant population in SAS cells, which comprise a large proportion of CSCs in oral cancer, that is, the CD24‑/CD44+ fraction. Furthermore, clinically relevant radioresistant cells (SAS‑R) are sensitive to Sal; therefore, further investigations are necessary.
View Figures

Figure 1

Measurements of the cancer stem cell
markers, CD24 and CD44. (A) Representative cytograms of SAS and
SAS-R cells. The fractions of (B) CD24+/CD44+
and (C) CD24-/CD44+ were measured from the
fractions shown in the cytograms. Values are presented as the mean
± standard deviation. **P<0.01, significant
differences.

Figure 2

Cell surviving fraction and sphere
formation following treatment with Sal. (A) The cell surviving
fraction of the SAS and SAS-R cells, Sal concentration (i.e., 0,
0.01, 0.03, 0.1, 0.3, 0.6 and 1 µM) gradient. Exp represents
experimental data. Blue and Red solid lines indicate log-logistic
sigmoidal curve fitting SAS and SAS-R cell surviving fraction,
respectively. (B) Representative images are shown. White scale
bars, 100 µm. (C) Sphere counts following treatment with 0.3 µM
Sal. Values are presented as the mean ± standard deviation.
**P<0.01, significant differences. Sal,
salinomycin.

Figure 3

Surviving fractions of SAS and SAS-R
cells following 2 Gy irradiation under Sal administration. These
surviving fractions were normalized to the respective untreated
controls for SAS (blue) and SAS-R (red) cells. Exp represents
experimental data.

Figure 4

Cell surviving fractions of SAS cells
and their subpopulations. (A) Representative colony images are
shown. The numbers indicate the number of seeded cells. (B)
Surviving fractions of the parental SAS cells and their
subpopulations (i.e., CD24+/CD44+ and
CD24-/CD44+ fractions). Values are presented
as the mean ± standard deviation. **P<0.01,
##P<0.01 and $$P<0.01, significant
differences between brackets (above the bars), DMSO group and 2 Gy
group, respectively. Sal, salinomycin.

Figure 5

Apoptotic cells treated with Sal
and/or 2 Gy irradiation. (A) Representative cytograms of apoptotic
cells 24 h following treatment with Sal and/or 2 Gy irradiation.
(B) Annexin V-positive fractions were defined as the apoptotic
fractions. Values are presented as the mean ± standard deviation.
*P<0.05 and **P<0.01, significant
differences. Sal, salinomycin.
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Spandidos Publications style
Saga R, Hosokawa Y and Fukumoto M: Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction. Med Int 6: 51, 2026.
APA
Saga, R., Hosokawa, Y., & Fukumoto, M. (2026). Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction. Medicine International, 6, 51. https://doi.org/10.3892/mi.2026.335
MLA
Saga, R., Hosokawa, Y., Fukumoto, M."Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction". Medicine International 6.5 (2026): 51.
Chicago
Saga, R., Hosokawa, Y., Fukumoto, M."Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction". Medicine International 6, no. 5 (2026): 51. https://doi.org/10.3892/mi.2026.335
Copy and paste a formatted citation
x
Spandidos Publications style
Saga R, Hosokawa Y and Fukumoto M: Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction. Med Int 6: 51, 2026.
APA
Saga, R., Hosokawa, Y., & Fukumoto, M. (2026). Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction. Medicine International, 6, 51. https://doi.org/10.3892/mi.2026.335
MLA
Saga, R., Hosokawa, Y., Fukumoto, M."Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction". Medicine International 6.5 (2026): 51.
Chicago
Saga, R., Hosokawa, Y., Fukumoto, M."Salinomycin is effective for clinically relevant radioresistant oral squamous cell carcinoma cancer cells and their cancer stem cell marker positive fraction". Medicine International 6, no. 5 (2026): 51. https://doi.org/10.3892/mi.2026.335
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