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Article

TGFβ1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells

  • Authors:
    • Nur Amilia Hanie Mohamad Hasan
    • Yu Zhao Lee
    • Chau Ling Tham
    • Daud Ahmad Israf
    • Nuzul Noorahya Jambari
    • Hanis Hazeera Harith
  • View Affiliations / Copyright

    Affiliations: Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia, School of Healthy Aging, Aesthetic and Regenerative Medicine, Faculty of Medicine and Health Sciences, UCSI University, Kuala Lumpur 56000, Malaysia, Department of Food Science, Faculty of Food and Science Technology, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
  • Article Number: 319
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    Published online on: September 15, 2025
       https://doi.org/10.3892/mmr.2025.13684
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Abstract

Epithelial‑mesenchymal transition (EMT) is a pathophysiological process contributing to bronchial remodeling in airway diseases such as chronic obstructive pulmonary disorder. EMT in several types of cancer involves dysregulated microtubule dynamics. Stathmin, a microtubule destabilizer, is highly expressed in different types of cancer, and is associated with decreased microtubule stability and enhanced migratory capability. The present study examined the relationship between stathmin expression and microtubule stability in bronchial EMT using an in vitro model. Primary normal human bronchial epithelial (NHBE) cells and the BEAS‑2B bronchial epithelial cell line were induced with TGFβ1 for 48 or 72 h to activate EMT, with or without the TGFβ1 inhibitor, SB431542. TGFβ1‑induced cells exhibited significantly reduced E‑cadherin (epithelial marker) and increased vimentin (mesenchymal marker) expression, which was inhibited by SB431542. TGFβ1‑mediated EMT was associated with reduced stathmin levels and increased microtubule stability (indicated by acetylated‑α‑tubulin) in BEAS‑2B and NHBE cells. However, TGFβ1‑induced EMT did not significantly enhance cell migration, potentially due to stabilized microtubules. By contrast, 10% fetal bovine serum induced a more robust EMT phenotype, accompanied by increased stathmin expression, reduced microtubule stability and enhanced cell migration. The present study highlights the potential role of stathmin in modulating microtubule dynamics during bronchial remodeling and hypothesizes its involvement in the transition from partial to full EMT, depending on the EMT‑inducing stimulus.
View Figures

Figure 1

Effects of TGFβ1-induced EMT on
stathmin and ace-α-tubulin expression in immortalized human
bronchial epithelial cells in the absence and presence of the TGFβ1
inhibitor SB431542 at 48 h. (A) Representative western blotting
images of stathmin, ace-α-tubulin, E-cadherin, vimentin and
α-tubulin following EMT induction by 10 ng/ml TGFβ1 with or without
the TGFβ1 inhibitor, SB431542 in BEAS-2B cells (from three
independent experiments). Densitometric analysis of (B) stathmin,
(C) ace-α-tubulin, (D) E-cadherin and (E) vimentin, normalized to
α-tubulin protein expression. Data are presented as the mean ± SEM
from three independent experiments; *P<0.05, **P<0.01,
***P<0.001; ns, not significant, using one-way ANOVA followed by
Tukey's post hoc test. Ace, acetylated; EMT, epithelial-mesenchymal
transition.

Figure 2

Effects of TGFβ1-induced EMT on
stathmin and ace-α-tubulin expression in immortalized human
bronchial epithelial cells in the absence and presence of the TGFβ1
inhibitor SB431542 at 72 h. (A) Representative western blotting
images of stathmin, ace-α-tubulin, E-cadherin, vimentin and
α-tubulin following EMT induction by 10 ng/ml TGFβ1 in BEAS-2B
cells. Densitometric analysis of (B) stathmin, (C) ace-α-tubulin,
(D) E-cadherin and (E) vimentin, normalized to α-tubulin protein
expression (from three independent experiments). Data are presented
as the mean ± SEM from three independent experiments; *P<0.05,
**P<0.01, ***P<0.001; ns, not significant, using one-way
ANOVA followed by Tukey's post hoc test. Ace, acetylated; EMT,
epithelial-mesenchymal transition.

Figure 3

Effects of TGFβ1-induced EMT on
stathmin and ace-α-tubulin expression in primary NHBE cells in the
absence and presence of the TGFβ1 inhibitor SB431542.
Representative western blotting images of stathmin, ace-α-tubulin,
E-cadherin, vimentin and α-tubulin following EMT induction by 10
ng/ml TGFβ1 in NHBE cells at (A) 48 h and (F) 72 h (from three
independent experiments). Densitometric analysis of the expression
of stathmin at (B) 48 and (G) 72 h, ace-α-tubulin at (C) 48 and (H)
72 h, E-cadherin at (D) 48 and (I) 72 h, and vimentin at (E) 48 and
(J) 72 h, normalized to α-tubulin. Data are presented as the mean ±
SEM from at least three independent experiments; *P<0.05,
**P<0.01, ***P<0.001, ****P<0.0001; ns, not significant,
using one-way ANOVA followed by Tukey's post hoc test. Ace,
acetylated; NHBE, normal human bronchial epithelial; EMT,
epithelial-mesenchymal transition.

Figure 4

Basal expression of stathmin,
ace-α-tubulin, E-cadherin and vimentin in primary bronchial
epithelial cells from individuals with COPD compared with NHBE
cells. (A) Representative western blotting images of stathmin,
ace-α-tubulin, E-cadherin, vimentin and α-tubulin in NHBE and COPD
bronchial epithelial cells at baseline (from three independent
experiments). Densitometric analysis of (B) stathmin, (C)
ace-α-tubulin, (D) E-cadherin and (E) vimentin expression,
normalized to α-tubulin expression. Data are presented as the mean
± SEM from three independent experiments; **P<0.01,
***P<0.001 and ****P<0.0001, using unpaired Student's t-test.
Ace, acetylated; NHBE, normal human bronchial epithelial; COPD,
chronic obstructive pulmonary disorder.

Figure 5

TGFβ1 induction does not alter the
migratory capability of human bronchial epithelial cells. (A)
Representative images of the scratch assay to assess migration of
BEAS-2B cells following TGFβ1 (10 ng/ml) induction in the absence
and presence of the TGFβ1 inhibitor SB431542 for 48 h and (B)
corresponding percentage of wound closure. (C) Representative
scratch assay images for 72 h and (D) corresponding percentage of
wound closure. Data are presented as the mean ± SEM from three
independent experiments; ns, not significant, using one-way ANOVA
followed by Tukey's post hoc test.

Figure 6

Differential regulation of stathmin
expression, microtubule stability and migratory phenotype in
bronchial epithelial cells following EMT induction by TGFβ1 or FBS
(10%) for 48 h. (A) Representative scratch assay images of BEAS-2B
cells following EMT induction by 10 ng/ml TGFβ1 (with or without
TGFβ1 inhibitor, SB431542) or 10% FBS and (B) corresponding
percentage of wound closure. (C) Representative western blotting
images, and densitometric analysis of (D) stathmin, (E)
ace-α-tubulin, (F) E-cadherin and (G) vimentin protein expression,
normalized to α-tubulin. Data are presented as the mean ± SEM from
at least three independent experiments; *P<0.05, **P<0.01,
***P<0.001, ****P<0.0001 vs. control group; ns, not
significant, using one-way ANOVA followed by Dunnett's post hoc
test. EMT, epithelial-mesenchymal transition; Ace, acetylated.

Figure 7

Differential regulation of stathmin
expression, microtubule stability and migratory phenotype in
bronchial epithelial cells following EMT induction by TGFβ1 or FBS
(10%) for 72 h. (A) Representative scratch assay images of BEAS-2B
cells following EMT induction by 10 ng/ml TGFβ1 (with or without
TGFβ1 inhibitor, SB431542) or 10% FBS and (B) corresponding
percentage of wound closure. (C) Representative western blotting
images, and the densitometric analysis of (D) stathmin, (E)
ace-α-tubulin, (F) E-cadherin and (G) vimentin protein expression,
normalized to α-tubulin. Data are presented as the mean ± SEM from
at least three independent experiments; *P<0.05, **P<0.01,
***P<0.001, ****P<0.0001 vs. control group; ns, not
significant, using one-way ANOVA followed by Dunnett's post hoc
test. EMT, epithelial-mesenchymal transition; Ace, acetylated.
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Copy and paste a formatted citation
Spandidos Publications style
Mohamad Hasan NH, Lee YZ, Tham CL, Israf DA, Jambari NN and Harith HH: TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells. Mol Med Rep 32: 319, 2025.
APA
Mohamad Hasan, N.H., Lee, Y.Z., Tham, C.L., Israf, D.A., Jambari, N.N., & Harith, H.H. (2025). TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells. Molecular Medicine Reports, 32, 319. https://doi.org/10.3892/mmr.2025.13684
MLA
Mohamad Hasan, N. H., Lee, Y. Z., Tham, C. L., Israf, D. A., Jambari, N. N., Harith, H. H."TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells". Molecular Medicine Reports 32.6 (2025): 319.
Chicago
Mohamad Hasan, N. H., Lee, Y. Z., Tham, C. L., Israf, D. A., Jambari, N. N., Harith, H. H."TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells". Molecular Medicine Reports 32, no. 6 (2025): 319. https://doi.org/10.3892/mmr.2025.13684
Copy and paste a formatted citation
x
Spandidos Publications style
Mohamad Hasan NH, Lee YZ, Tham CL, Israf DA, Jambari NN and Harith HH: TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells. Mol Med Rep 32: 319, 2025.
APA
Mohamad Hasan, N.H., Lee, Y.Z., Tham, C.L., Israf, D.A., Jambari, N.N., & Harith, H.H. (2025). TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells. Molecular Medicine Reports, 32, 319. https://doi.org/10.3892/mmr.2025.13684
MLA
Mohamad Hasan, N. H., Lee, Y. Z., Tham, C. L., Israf, D. A., Jambari, N. N., Harith, H. H."TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells". Molecular Medicine Reports 32.6 (2025): 319.
Chicago
Mohamad Hasan, N. H., Lee, Y. Z., Tham, C. L., Israf, D. A., Jambari, N. N., Harith, H. H."TGF&beta;1‑induced epithelial‑mesenchymal transition is associated with stathmin downregulation and increased microtubule stability in bronchial epithelial cells". Molecular Medicine Reports 32, no. 6 (2025): 319. https://doi.org/10.3892/mmr.2025.13684
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