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Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest

  • Authors:
    • Minji Bae
    • Min Kyeong Lee
    • Mi Gyeong Jeong
    • Yeonseo Lee
    • Harim Joo
    • Young Chan Lee
    • Jung Woo Lee
    • Su Il Kim
    • Seong-Gyu Ko
    • Young-Gyu Eun
  • View Affiliations / Copyright

    Affiliations: Department of Biomedical Science and Technology, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea, Department of Otolaryngology‑Head and Neck Surgery, Kyung Hee University School of Medicine, Kyung Hee University Medical Center, Seoul 02447, Republic of Korea, Department of Oral and Maxillofacial Surgery, School of Dentistry, Kyung Hee University, Seoul 02447, Republic of Korea, Department of Preventive Medicine, College of Korean Medicine, Kyung Hee University, Seoul 02447, Republic of Korea
    Copyright: © Bae et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 83
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    Published online on: February 27, 2026
       https://doi.org/10.3892/or.2026.9088
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Abstract

Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignancies worldwide. Cisplatin, a widely used chemotherapeutic agent, exerts its anticancer effect by inducing DNA damage, and inhibiting transcription and replication. However, its repeated administration can lead to severe side effects and the development of drug resistance, thereby limiting its clinical efficacy. Thus, novel combination strategies are urgently needed to enhance the anticancer effects of cisplatin. The present study aimed to investigate the potential of Platycodin D (PD), a natural saponin extracted from Platycodon grandiflorus, to enhance cisplatin sensitivity in HNSCC cells. The results demonstrated that combination treatment with PD and cisplatin synergistically reduced cell viability and markedly suppressed colony formation compared with either agent alone. Furthermore, the combination treatment markedly increased intracellular reactive oxygen species (ROS) levels, and markedly downregulated the expression levels of antioxidant genes, including heme oxygenase‑1, NAD(P)H quinone dehydrogenase 1, superoxide dismutase 1 and sulfiredoxin 1. Additionally, the combination treatment excessively activated autophagy, whereas PD inhibited autophagic flux, as determined by LC3A/B and p62 accumulation following Bafilomycin A1 treatment, ultimately promoting apoptosis. These findings suggested that PD may serve as a potential cisplatin sensitizer by enhancing ROS accumulation and disrupting autophagy, thereby presenting a promising combination therapeutic strategy for the treatment of HNSCC.
View Figures

Figure 1

PD and cisplatin combination
treatment synergistically reduces viability in head and neck
squamous cell carcinoma cells. (A) HSC3 and FaDu cells were treated
with PD (5, 10 or 15 µM), cisplatin (5 or 10 µM) or combination
treatment for 48 h. Cell viability was assessed using an MTT assay.
Statistical significance was determined by one-way ANOVA. (B) CI
values were calculated, with a CI <1 indicating a synergistic
effect. Fa (fraction affected) represents the fraction of cells
affected by drug treatment and was calculated based on cell
viability data. Data are presented as the mean ± SD (n≥3).
*P<0.05, **P<0.01, ***P<0.001. PD, Platycodin D; CI,
combination index; Fa, fraction affected.

Figure 2

Combination of PD and cisplatin
markedly suppresses colony formation in head and neck squamous cell
carcinoma cells. (A) HSC3 and (B) FaDu cells were seeded at 300
cells per well and treated with PD (5 µM), cisplatin (5 µM) or
combination treatment for 7–10 days. The quantitative graph
represents the number of colonies. Statistical significance was
determined by one-way ANOVA followed by Tukey's post hoc test. Data
are presented as the mean ± SD (n=3). **P<0.01, ***P<0.001.
PD, Platycodin D.

Figure 3

Combination of PD and cisplatin
downregulates antioxidant gene expression in head and neck squamous
cell carcinoma cells. (A) HSC3 and (B) FaDu cells were treated with
PD (10 µM), cisplatin (10 µM) or combination treatment for 12 h
(HSC3 cells) or 3 h (FaDu cells). mRNA expression levels of
antioxidant genes (HO-1, NQO1, SOD1 and SRXN1) were measured by
reverse transcription-quantitative PCR and normalized to β-actin
expression. Combination treatment markedly reduced the expression
of all four genes compared with single treatments. Statistical
significance was determined by one-way ANOVA followed by Tukey's
post hoc test. *P<0.05, **P<0.01, ***P<0.001. PD,
Platycodin D; HO-1, heme oxygenase-1; NQO1, NAD(P)H quinone
dehydrogenase 1; SOD1, superoxide dismutase 1; SRXN1, sulfiredoxin
1.

Figure 4

Combination of PD and cisplatin
synergistically increases ROS levels in head and neck squamous cell
carcinoma cells. (A) HSC3 and (B) FaDu cells were treated with PD
(10 µM), cisplatin (10 µM) or combination treatment, and
intracellular ROS levels were assessed by DCF-DA staining.
Combination treatment markedly increased ROS levels compared with
single treatments. Statistical significance was determined by
one-way ANOVA followed by Tukey's post hoc test. In the graphs,
vehicle, PD, cisplatin and combination treatment groups are
represented by black, green, blue and red lines, respectively. FL
indicates fluorescence intensity measured by flow cytometry and
reflects intracellular ROS levels. *P<0.05, **P<0.01,
***P<0.001. PD, Platycodin D; ROS, reactive oxygen species;
DCF-DA, 2′,7′-dichlorodihydrofluorescein diacetate.

Figure 5

PD and cisplatin combination
treatment induces morphological changes and vesicle accumulation.
(A) HSC3 and (B) FaDu cells were treated with PD (10 µM), cisplatin
(10 µM) or combination treatment for 48 h. Cell morphology was
observed under a light microscope to evaluate vesicular structure
accumulation. The images on the right show enlarged views of the
boxed regions in the images on the left. Increased vesicular
structures were observed in cells treated with PD alone or in
combination with cisplatin and arrows indicate representative
vesicular structures. Scale bar, 500 µm. PD, Platycodin D.

Figure 6

PD and cisplatin combination
treatment increases autophagy marker levels in head and neck
squamous cell carcinoma cells. (A) HSC3 and (C) FaDu cells were
treated with PD (10 µM), cisplatin (10 µM) or combination treatment
for 48 h. Cells were stained with LC3B antibody and analyzed using
confocal microscopy. (B) In HSC3 cells, p62 immunofluorescence was
increased following combination treatment compared with single
treatments. (D) In FaDu cells, p62 staining similarly showed higher
p62 levels in the combination treatment group than in the single
treatment groups. Nuclei were stained with DAPI (blue), LC3B was
stained red and p62 was stained green. Fluorescence intensity and
puncta quantification were analyzed using ZEN software (Zeiss AG).
Scale bar, 50 µm. Data are presented as the mean ± SD (n=3).
*P<0.05, **P<0.01, ***P<0.001. PD, Platycodin D.

Figure 7

PD and cisplatin combination
treatment induces autophagy arrest by inhibiting autophagosome
degradation. HSC3 and FaDu cells were treated with PD (10 µM),
cisplatin (10 µM) or combination treatment for 48 h, with or
without BafA1. (A) Western blotting was performed to evaluate the
expression levels of LC3A/B and p62. β-actin was used as a loading
control. (B) Densitometric analysis of LC3A/B and p62 protein
levels normalized to β-actin, performed using ImageJ software.
Statistical significance was determined by one-way ANOVA followed
by Tukey's post hoc test. Data are presented as the mean ± SD
(n=3). *P<0.05, **P<0.01. BafA1, Bafilomycin A1; PD,
Platycodin D.

Figure 8

PD and cisplatin combination
treatment induces apoptotic cell death in head and neck squamous
cell carcinoma cells. (A) Apoptosis was analyzed by annexin V/PI
staining followed by flow cytometric analysis in HSC3 and FaDu
cells treated with PD (10 µM), cisplatin (10 µM) or their
combination for 48 h. Combination treatment markedly increased the
percentage of apoptotic cells. Data are presented as the mean ± SD
(n=3). *P<0.05, **P<0.01, ***P<0.001. (B) Western blot
analysis was performed to evaluate the levels of apoptosis-related
proteins PARP and cleaved-caspase 3. β-actin was used as a loading
control. PD, Platycodin D; PARP, poly(ADP-ribose) polymerase.
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Copy and paste a formatted citation
Spandidos Publications style
Bae M, Lee MK, Jeong MG, Lee Y, Joo H, Lee YC, Lee JW, Kim SI, Ko S, Eun Y, Eun Y, et al: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest. Oncol Rep 55: 83, 2026.
APA
Bae, M., Lee, M.K., Jeong, M.G., Lee, Y., Joo, H., Lee, Y.C. ... Eun, Y. (2026). Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest. Oncology Reports, 55, 83. https://doi.org/10.3892/or.2026.9088
MLA
Bae, M., Lee, M. K., Jeong, M. G., Lee, Y., Joo, H., Lee, Y. C., Lee, J. W., Kim, S. I., Ko, S., Eun, Y."Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest". Oncology Reports 55.5 (2026): 83.
Chicago
Bae, M., Lee, M. K., Jeong, M. G., Lee, Y., Joo, H., Lee, Y. C., Lee, J. W., Kim, S. I., Ko, S., Eun, Y."Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest". Oncology Reports 55, no. 5 (2026): 83. https://doi.org/10.3892/or.2026.9088
Copy and paste a formatted citation
x
Spandidos Publications style
Bae M, Lee MK, Jeong MG, Lee Y, Joo H, Lee YC, Lee JW, Kim SI, Ko S, Eun Y, Eun Y, et al: Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest. Oncol Rep 55: 83, 2026.
APA
Bae, M., Lee, M.K., Jeong, M.G., Lee, Y., Joo, H., Lee, Y.C. ... Eun, Y. (2026). Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest. Oncology Reports, 55, 83. https://doi.org/10.3892/or.2026.9088
MLA
Bae, M., Lee, M. K., Jeong, M. G., Lee, Y., Joo, H., Lee, Y. C., Lee, J. W., Kim, S. I., Ko, S., Eun, Y."Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest". Oncology Reports 55.5 (2026): 83.
Chicago
Bae, M., Lee, M. K., Jeong, M. G., Lee, Y., Joo, H., Lee, Y. C., Lee, J. W., Kim, S. I., Ko, S., Eun, Y."Platycodin D sensitizes head and neck squamous cell carcinoma to cisplatin by inducing autophagy arrest". Oncology Reports 55, no. 5 (2026): 83. https://doi.org/10.3892/or.2026.9088
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