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Article

TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells

  • Authors:
    • Maki Nara
    • Yoshiki Mukudai
    • Masataka Watanabe
    • Nodoka Kindaichi
    • Konomi Yamada
    • Yuzo Abe
    • Toshikazu Shimane
    • Tatsuo Shirota
    • Seigo Ohba
  • View Affiliations / Copyright

    Affiliations: Department of Oral and Maxillofacial Surgery, School of Dentistry, Showa Medical University, Tokyo 145‑8515, Japan
  • Article Number: 164
    |
    Published online on: July 24, 2026
       https://doi.org/10.3892/or.2026.9169
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Abstract

Tumor protein D52 (TPD52) family proteins are involved in the proliferation, survival and malignant progression of oral squamous cell carcinoma (OSCC). However, their roles in chemoresistance remain incompletely understood. The present study investigated the contribution of TPD52 family proteins to anticancer drug resistance, with particular emphasis on tumor protein D54 (TPD54). OSCC cells were treated with cisplatin, 5‑fluorouracil, or docetaxel (DTX), and the expression of TPD52 family members was examined. Gain‑ and loss‑of‑function analyses were performed to evaluate cell viability, apoptotic responses, cytochrome p450 (P450) and P‑glycoprotein (P‑gp) activities, protein expression, intracellular localization and membrane/cytosol distribution. Anticancer drug treatment increased the expression of TPD52, TPD53 and TPD54. Among these family members, TPD54 showed the strongest association with DTX resistance by attenuating the reduction in cell viability without affecting cell‑cycle progression. TPD54 overexpression attenuated DTX‑associated apoptotic responses and was associated with changes in apoptosis‑, ferroptosis‑, and autophagy‑related marker proteins. TPD54 expression had little effect on the activities of P450 3A4 or P450 1B1 but significantly increased P‑gp activity. Membrane/cytosol fractionation demonstrated increased membrane localization of endogenous P‑gp following TPD54 overexpression, whereas co‑immunoprecipitation and immunocytofluorescence analyses revealed an association and partial co‑localization between TPD54 and P‑gp. These findings suggest that TPD54 contributes to DTX resistance in OSCC cells through modulation of P‑gp localization and activity. The present study identifies TPD54 as a potential contributor to P‑gp‑associated chemoresistance and provides a basis for further investigation of the molecular mechanisms underlying multidrug resistance in OSCC.
View Figures

Figure 1

Effects of CDDP, 5-FU, and DTX on the
mRNA expression of TPD52 family members. SAS cells were exposed to
CDDP or 5-FU (0, 1, and 10 µM) or DTX (0, 0.1, and 1 µM) for 48 h.
The mRNA expression of TPD52, TPD53, and TPD54 was analyzed by
RT-qPCR. Data are presented as the mean ± SD of three independent
experiments. *P<0.05 vs. control (0 µM). CDDP, cisplatin; 5-FU,
5-fluorouracil; DTX, docetaxel; RT-qPCR, reverse
transcription-quantitative polymerase chain reaction.

Figure 2

Effects of overexpression or knockdown
of TPD52 family proteins on cell viability. Overexpression vectors
or small interfering RNAs for TPD52, TPD53 and TPD54 were
transfected into SAS cells. Cells were incubated in the absence (−)
or presence of CDDP, 5-FU, or DTX for 48 h, and cell viability was
assessed using the MTT assay. Data are presented as the mean ± SD
of three independent experiments. *P<0.05 vs. untreated control.
CDDP, cisplatin; 5-FU, 5-fluorouracil; DTX, docetaxel; MTT,
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

Figure 3

Effects of overexpression or knockdown
of TPD52 family proteins on cell-cycle progression. (A)
Overexpression vectors or (B) siRNAs were transfected into SAS
cells. Cells were incubated in the absence (−) or presence of CDDP,
5-FU, or DTX for 48 h, and cell-cycle distribution was analyzed.
Blue, G0/G1; orange, G1; gray, S; yellow, G2/M. CDDP, cisplatin;
5-FU, 5-fluorouracil; DTX, docetaxel.

Figure 4

Effects of overexpression or knockdown
of TPD52 family proteins on apoptotic responses and cell
death-related marker proteins. Overexpression vectors or siRNAs for
TPD52, TPD53, and TPD54 were transfected into SAS cells. Cells were
incubated in the absence (−) or presence (+) of CDDP, 5-FU, or DTX
for 48 h. (A and B) Caspase-3/7 activity and western blot analysis
of LC3, p62, Bax, Bcl-2, GPX4 and xCT/SLC7A11 were performed.
Representative western blots from three independent experiments are
shown. *P<0.05 vs. untreated control. CDDP, cisplatin; 5-FU,
5-fluorouracil; DTX, docetaxel.

Figure 5

Effects of overexpression or knockdown
of TPD54 on DTX-associated apoptotic responses in SAS cells.
Overexpression vectors or siRNAs for TPD54 were transfected into
SAS cells. (A-C) Cells were incubated in the absence (−) or
presence (+) of DTX for 48 h, followed by assays for (A)
caspase-3/7 activity, (B) caspase-9 activity and (C) mitochondrial
membrane potential. Data are presented as the mean ± SD of three
independent experiments. *P<0.05. DTX, docetaxel.

Figure 6

Effects of overexpression or knockdown
of TPD54 on the expression of cell death-related proteins in SAS
cells. Overexpression vectors (HaloTag or HaloTag-TPD54) or small
interfering RNAs were transfected into SAS cells. Cells were
incubated in the absence (−) or presence (+) of DTX for 48 h and
subjected to western blot analysis for HaloTag, TPD54, P-gp, Bax,
Bcl-2, GPX4, xCT/SLC7A11 and GAPDH. Representative western blots
from three independent experiments are shown. P-gp, P-glycoprotein;
GPX4, glutathione peroxidase 4; DTX, docetaxel.

Figure 7

Effects of overexpression or knockdown
of TPD54 on the expression of cell death-related proteins in HSC-3
cells. Overexpression vectors (HaloTag or HaloTag-TPD54) or small
interfering RNAs were transfected into HSC-3 cells. Cells were
incubated in the absence (−) or presence (+) of DTX for 48 h and
subjected to western blot analysis for HaloTag, TPD54, P-gp, Bax,
Bcl-2, GPX4, xCT/SLC7A11 and GAPDH. Representative western blots
from three independent experiments are shown. P-gp, P-glycoprotein;
GPX4, glutathione peroxidase 4; DTX, docetaxel.

Figure 8

Effects of overexpression or knockdown
of TPD54 on the activities of P450 enzymes and P-gp. (A-C) SAS
cells were transfected and reseeded as described for the MTT assay.
The activities of (A) P450 3A4, (B) P450 1B1 and (C) P-gp were
measured. P-gp activity was evaluated under control conditions or
in the presence of CsA or VER. Data are presented as the mean ± SD
of three independent experiments. P-gp, P-glycoprotein; CsA,
cyclosporin A; VER, verapamil; DTX, docetaxel.

Figure 9

Association of TPD54 with
intracellular localization of P-gp. (A) HaloTag
co-immunoprecipitation assay. HaloTag or HaloTag-TPD54 expression
vectors were transfected into SAS cells, followed by HaloTag
immunoprecipitation and western blot analysis for HaloTag, TPD54,
P-gp, TPD53, E-cadherin and GAPDH. (B) Immunocytofluorescence
analysis of endogenous TPD53, TPD54 and P-gp in SAS cells with DAPI
nuclear staining. Representative results from three independent
experiments are shown. P-gp, P-glycoprotein; DTX, docetaxel.
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Copy and paste a formatted citation
Spandidos Publications style
Nara M, Mukudai Y, Watanabe M, Kindaichi N, Yamada K, Abe Y, Shimane T, Shirota T and Ohba S: TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells. Oncol Rep 56: 164, 2026.
APA
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y. ... Ohba, S. (2026). TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells. Oncology Reports, 56, 164. https://doi.org/10.3892/or.2026.9169
MLA
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y., Shimane, T., Shirota, T., Ohba, S."TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells". Oncology Reports 56.4 (2026): 164.
Chicago
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y., Shimane, T., Shirota, T., Ohba, S."TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells". Oncology Reports 56, no. 4 (2026): 164. https://doi.org/10.3892/or.2026.9169
Copy and paste a formatted citation
x
Spandidos Publications style
Nara M, Mukudai Y, Watanabe M, Kindaichi N, Yamada K, Abe Y, Shimane T, Shirota T and Ohba S: TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells. Oncol Rep 56: 164, 2026.
APA
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y. ... Ohba, S. (2026). TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells. Oncology Reports, 56, 164. https://doi.org/10.3892/or.2026.9169
MLA
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y., Shimane, T., Shirota, T., Ohba, S."TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells". Oncology Reports 56.4 (2026): 164.
Chicago
Nara, M., Mukudai, Y., Watanabe, M., Kindaichi, N., Yamada, K., Abe, Y., Shimane, T., Shirota, T., Ohba, S."TPD54 contributes to docetaxel resistance through modulation of P‑glycoprotein localization and activity in oral squamous cell carcinoma cells". Oncology Reports 56, no. 4 (2026): 164. https://doi.org/10.3892/or.2026.9169
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