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miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5

  • Authors:
    • Hiroto Tanaka
    • Suguru Maruyama
    • Katsutoshi Shoda
    • Yoshihiko Kawaguchi
    • Yudai Higuchi
    • Takaomi Ozawa
    • Takashi Nakayama
    • Ryo Saito
    • Wataru Izumo
    • Koichi Takiguchi
    • Kensuke Shiraishi
    • Shinji Furuya
    • Hidetake Amemiya
    • Hiromichi Kawaida
    • Daisuke Ichikawa
  • View Affiliations / Copyright

    Affiliations: Department of Digestive Surgery, University of Yamanashi, Chuo, Yamanashi 409‑3898, Japan
    Copyright: © Tanaka et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 170
    |
    Published online on: October 3, 2025
       https://doi.org/10.3892/or.2025.9003
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Abstract

Poorly differentiated cancers, including esophageal squamous cell carcinoma (ESCC), exhibit higher malignant potential and worse prognoses than well‑differentiated types. The present study aimed to identify microRNAs (miRNAs or miRs) involved in ESCC progression and their target mRNAs, focusing on tumor differentiation. miRNA candidates were selected using a miRNA array‑based approach and GEO datasets, comparing expression levels between poorly and non‑poorly differentiated ESCC. Clinical samples (n=61) and cell lines were analyzed to determine the significance and function of the selected miRNAs and their target mRNA. miR‑100‑5p and miR‑203a‑3p were significantly downregulated in poorly differentiated ESCC, with lower expression strongly associated with poorer overall survival (OS) (miR‑100‑5p: P=0.02; miR‑203a‑3p: P=0.05) and relapse‑free survival (RFS) (miR‑100‑5p: P=0.04; miR‑203a‑3p: P=0.12). Overexpression of these miRNAs suppressed cell migration and invasion. FKBP5 was identified as a common target, with its expression significantly reduced upon double‑transfection with miR‑100‑5p and miR‑203a‑3p. FKBP5 downregulation reduced tumor aggressiveness in KYSE70 cells, and clinical samples showed significantly worse survival rates in patients with high FKBP5 expression (OS: P=0.02; RFS: P=0.04). These findings suggest that miR‑100‑5p and miR‑203a‑3p act as tumor suppressors by targeting FKBP5, highlighting FKBP5 as a potential therapeutic target in ESCC.
View Figures

Figure 1

(A) miRNA expression ratio in non-por
portion relative to por portion of the tumor using global
normalization. (B) Venn diagram showing miRNAs more highly
expressed in non-por than por in both microarray and GEO datasets.
miRNA or miR, microRNA; por, poorly differentiated; non-por, well-
to moderately differentiated.

Figure 2

(A) Relationship between tumor
differentiation and miR-100-5p and miR-203a-3p levels in clinical
samples. (B and C) Kaplan-Meier curves for overall survival and
relapse-free survival rates in patients stratified by (B)
miR-100-5p levels and (C) miR-203a-3p levels. All the quantitative
measurements were performed in triplicate, and data for relative
quantity are presented as the mean ± SD. *P<0.05. por, poorly
differentiated. non-por, well- to moderately differentiated.

Figure 3

(A) Relationship between tumor
differentiation and miR-100-5p and miR-203a-3p levels in non-por
and por cell lines. Representative images and quantitative
measurement of (B and C) migration and (D and E) invasion assays
induced by miR-100-5p and miR-203a-3p mimics in KYSE70 cells. For
double-transfection experiments, both miR-100-5p and miR-203a-3p
mimics were co-transfected at 10 nM each. Scale bar, 200 µm. All
the quantitative measurements were performed in triplicate, and
data for relative quantity are presented as the mean ± SD.
**P<0.01, ***P<0.001 and ****P<0.0001 compared with NC.
miR, microRNA; por cell lines, poorly differentiated cell lines
(KYSE70, KYSE110, TE5) non-por cell lines, well- to moderately
differentiated cell lines (TE4, TE8, TE11). NC, negative
control.

Figure 4

Biological functions of the
FKBP5 gene. (A) Gene Ontology enrichment analysis of the
FKBP5 gene. (B) Kyoto Encyclopedia of Genes and Genomes
enrichment analysis of the FKBP5 gene.

Figure 5

Biological functions of the
FKBP5 gene. (A-D) Positive association between FKBP5
and (A) Vimentin, (B) N-cadherin, (C) PIK3 family and (D) AKT
family. (E and F) FKBP5 regulated (E) epithelial-mesenchymal
transition and (F) PI3K/AKT signaling pathway using the Gene Set
Enrichment Analysis.

Figure 6

FKBP5 expression in KYSE70
cells transfected with miR-100-5p and miR-203a-3p mimics. (A)
Relative mRNA expression levels measured by reverse
transcription-quantitative PCR. (B) Protein levels determined by
western blotting. **P<0.01 and ***P<0.001. miR, microRNA; NC,
negative control.

Figure 7

Representative images and
quantitative measurement of (A and B) migration, (C and D) invasion
and (E) cytotoxicity assays induced by FKBP5 suppression in
KYSE70 cells. Scale bar, 200 µm. All the quantitative measurements
were performed in triplicate, and data for relative quantity are
presented as the mean ± SD. *P<0.05 and ****P<0.0001 compared
with NC. NC, negative control; si-, small interfering.

Figure 8

Characterization of (A)
epithelial-to-mesenchymal transition markers (E-cadherin and
Vimentin) and (B) PI3K/ AKT signaling pathway components, including
p-PI3K, p-AKT and total (PI3K and AKT) proteins, in response to
FKBP5 regulation by western blotting in KYSE70 cells.
Quantification of (C) p-PI3K/ PI3K and (D) p-AKT/ AKT ratios
expressed as fold change relative to NC. *P<0.05. p-,
phosphorylated; NC, negative control; si-, small interfering.

Figure 9

(A) Representative
immunohistochemical staining for FKBP5 in esophageal
squamous cell carcinoma tissues by IHC. Scale bar, 20 µm. (B)
Kaplan-Meier curves for overall survival and relapse-free survival
rates in patients stratified by FKBP5.

Figure 10

Schematic representation of the
proposed role of miR-100-5p and miR-203a-3p in esophageal squamous
cell carcinoma. miR-100-5p and miR-203a-3p inhibit tumor
progression by targeting FKBP5. miRNA or miR, microRNA.
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Copy and paste a formatted citation
Spandidos Publications style
Tanaka H, Maruyama S, Shoda K, Kawaguchi Y, Higuchi Y, Ozawa T, Nakayama T, Saito R, Izumo W, Takiguchi K, Takiguchi K, et al: miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5. Oncol Rep 54: 170, 2025.
APA
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T. ... Ichikawa, D. (2025). miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5. Oncology Reports, 54, 170. https://doi.org/10.3892/or.2025.9003
MLA
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T., Nakayama, T., Saito, R., Izumo, W., Takiguchi, K., Shiraishi, K., Furuya, S., Amemiya, H., Kawaida, H., Ichikawa, D."miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5". Oncology Reports 54.6 (2025): 170.
Chicago
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T., Nakayama, T., Saito, R., Izumo, W., Takiguchi, K., Shiraishi, K., Furuya, S., Amemiya, H., Kawaida, H., Ichikawa, D."miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5". Oncology Reports 54, no. 6 (2025): 170. https://doi.org/10.3892/or.2025.9003
Copy and paste a formatted citation
x
Spandidos Publications style
Tanaka H, Maruyama S, Shoda K, Kawaguchi Y, Higuchi Y, Ozawa T, Nakayama T, Saito R, Izumo W, Takiguchi K, Takiguchi K, et al: miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5. Oncol Rep 54: 170, 2025.
APA
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T. ... Ichikawa, D. (2025). miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5. Oncology Reports, 54, 170. https://doi.org/10.3892/or.2025.9003
MLA
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T., Nakayama, T., Saito, R., Izumo, W., Takiguchi, K., Shiraishi, K., Furuya, S., Amemiya, H., Kawaida, H., Ichikawa, D."miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5". Oncology Reports 54.6 (2025): 170.
Chicago
Tanaka, H., Maruyama, S., Shoda, K., Kawaguchi, Y., Higuchi, Y., Ozawa, T., Nakayama, T., Saito, R., Izumo, W., Takiguchi, K., Shiraishi, K., Furuya, S., Amemiya, H., Kawaida, H., Ichikawa, D."miR‑100‑5p and miR‑203a‑3p suppress esophageal squamous cell carcinoma progression by targeting FKBP5". Oncology Reports 54, no. 6 (2025): 170. https://doi.org/10.3892/or.2025.9003
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