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Article Open Access

YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion

  • Authors:
    • Qiang Li
    • Yuan Wang
    • Zhao Yuan
    • Peng Zhai
    • Jian Wang
    • Chen Zhang
    • Ziqi Shao
    • Chungen Xing
  • View Affiliations / Copyright

    Affiliations: Department of General Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China, Department of General Surgery of the Second Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221000, P.R. China, Department of General Surgery, Huai'an Fifth People's Hospital, Huai'an, Jiangsu 223300, P.R. China, Department of General Surgery, The Affiliated Huaian First People's Hospital of Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 228
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    Published online on: October 20, 2025
       https://doi.org/10.3892/ijmm.2025.5669
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Abstract

The present study investigated the role of YWHAH in colorectal cancer (CRC) progression, focusing on its regulation of autophagy via the MAPK/ERK signaling pathway and subsequent effects on cell migration and invasion. Clinical sample analysis revealed that YWHAH was highly expressed in CRC tissues, associating with poor differentiation, advanced TNM stage, lymph node metastasis, vascular invasion and unfavorable prognosis. In vitro experiments showed that YWHAH overexpression promoted proliferation, and epithelial‑mesenchymal transition of CRC cells, while inhibiting apoptosis. Conversely, YWHAH knockdown exerted opposite effects. Autophagy assays demonstrated that YWHAH knockdown enhanced autophagy by upregulating autophagy related 7 and LC3II/I and downregulating P62, whereas overexpression suppressed autophagy. Mechanistically, YWHAH negatively regulated the MAPK/ERK pathway [reducing phosphorylated (p)‑ERK1/2 and p‑ELK‑1 levels] to inhibit autophagy. The ERK inhibitor PD98059 reversed autophagy activation by YWHAH knockdown, while the ERK agonist U‑46619 reversed autophagy suppression by YWHAH overexpression. Additionally, the autophagy inhibitor 3‑methyladenine abrogated the inhibitory effects of YWHAH knockdown on migration and invasion, and the autophagy inducer rapamycin reversed the promoting effects of YWHAH overexpression. In vivo, nude mouse xenograft models confirmed that YWHAH inhibited autophagy via the MAPK/ERK pathway to accelerate tumor growth. These findings underscore the role of YWHAH as a critical regulator of CRC progression and suggest it as a potential therapeutic target. Interventions targeting YWHAH or its downstream factors may provide innovative approaches for treating CRC, particularly by modulating autophagy to inhibit tumor growth and metastasis.
View Figures

Figure 1

Expression and prognostic value of
YWHAH. (A) The expression distribution of YWHAH in CRC and normal
tissues using R package. (B) Relative expression of YWHAH in CRC
tissue and PCT using reverse transcription-quantitative PCR. (C)
Analysis of survival rates in patients with CRC with high and low
expression of YWHAH. (D) Relative expression of YWHAH in colorectal
tumor and PCT using immunohistochemistry. *P<0.05 and
****P<0.0001. YWHAH, tyrosine
3-monooxygenase/tryptophan 5-monooxygenase activation protein Eta;
CRC, colorectal cancer; PCT, paracancerous tissue.

Figure 2

Impact of differential YWHAH
expression levels on colorectal cancer cell proliferation in
vitro and in vivo. (A and B) Cell Counting Kit-8 was
used to detect cell viability. (C and D) Impact of different YWHAH
expression levels on tumor formation in nude mice. (E and F) The
tumor volume was determined. *P<0.05,
**P<0.01 and ***P<0.001. Data are
presented as the mean ± standard deviation (n=3 independent
experiments). YWHAH, tyrosine 3-monooxygenase/tryptophan
5-monooxygenase activation protein Eta; OE, overexpression; si-,
small interfering; NC, negative control.

Figure 3

Western blot assay is used to detect
the effect of YWHAH on EMT in colorectal cancer cells. (A and B)
Western blot assay was used to detect the impact of YWHAH
overexpression and inhibition on EMT. (C and D) Semi-quantitative
analysis of EMT-related protein expression levels. Data are
presented as the mean ± standard deviation (n=3 independent
experiments). *P<0.05, **P<0.01 and
***P<0.0001. YWHAH, tyrosine
3-monooxygenase/tryptophan 5-monooxygenase activation protein Eta;
EMT, epithelial-to-mesenchymal transition; OE, overexpression; si-,
small interfering; NC, negative control.

Figure 4

Effect of YWHAH on apoptosis of
different CRC cells. (A) Apoptosis of LOVO cells was measured by
flow cytometry quantitative assay of apoptosis. (B) Apoptosis of
HCT116 cells was measured by flow cytometry quantitative assay of
apoptosis. (C and D) Western blot analysis of Caspase3, BAX and
Bcl2 protein expression in CRC cells. (E and F) Quantification of
Caspase3 protein expression. (G and H) Ratio of Bcl2 and BAX
protein. Data are presented as the mean ± standard deviation (n=3
independent experiments). *P<0.05,
**P<0.01, ***P<0.001 and
****P<0.0001. YWHAH, tyrosine
3-monooxygenase/tryptophan 5-monooxygenase activation protein Eta;
CRC, colorectal cancer; OE, overexpression; si-, small interfering;
NC, negative control.

Figure 5

Impact of different YWHAH expression
levels on autophagy in colorectal cancer cells. (A and B)
Monitoring of the double-tagged LC3 by RFP-GFP-hLC3 tandem
fluorescent protein in LOVO and HCT116 cells. (C and D)
Quantification of GFP−/RFP+/LC3 dots in LOVO
and HCT116 cells. (E and F) Western blot analysis of ATG7, P62 and
LC3II/LC3I protein expression. Data are presented as the mean ±
standard deviation (n=3 independent experiments).
*P<0.05, **P<0.01 and
***P<0.001. YWHAH, tyrosine
3-monooxygenase/tryptophan 5-monooxygenase activation protein Eta;
ATG7, autophagy related 7; OE, overexpression; si-, small
interfering; NC, negative control.

Figure 6

Inhibition of autophagy reverses the
suppressive effects of si-YWHAH on migration and invasion in CRC
cells. (A and B) Western blot assay detected the expression of
autophagy-related proteins in CRC cells after the addition of the
autophagy inhibitor 3-MA. (C and D) Quantitative analysis of
changes in migration of CRC cells following 3-MA treatment and
YWHAH knockdown. (E and F) Quantitative analysis of changes in
invasion of CRC cells following 3-MA treatment and YWHAH knockdown.
Data are presented as the mean ± standard deviation (n=3
independent experiments). *P<0.05,
**P<0.01 and ***P<0.001. YWHAH,
tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation
protein Eta; si-, small interfering; 3-MA, 3-methyladenine; CRC,
colorectal cancer; NC, negative control.

Figure 7

Activation of autophagy reverses the
promoting effect of OE-YWHAH on the migration and invasion of CRC
cells. (A and B) Western blot assay detected the expression of
autophagy-related proteins in CRC cells after the addition of the
autophagy activator RAPA. (C and D) Quantitative analysis of
changes in migration of CRC cells following RAPA treatment and
YWHAH overexpression. (E and F) Quantitative analysis of changes in
invasion of CRC cells following RAPA treatment and YWHAH
overexpression. Data are presented as the mean ± standard deviation
(n=3 independent experiments). *P<0.05,
**P<0.01 and ***P<0.001. OE-,
overexpression; YWHAH, tyrosine 3-monooxygenase/tryptophan
5-monooxygenase activation protein Eta; CRC, colorectal cancer;
RAPA, rapamycin; si-, small interfering; NC, negative control.

Figure 8

Western blot assay detects the
expression of autophagy-related proteins and MAPK/ERK signaling
pathway-related proteins in colorectal cancer cells after the
addition of the MAPK/ERK pathway inhibitor PD-98059. (A and B)
Western blot assays was sued to detect the expression of
autophagy-related proteins and MAPK/ERK signaling pathway-related
proteins. (C-F) Semi-quantitative analysis of autophagy-related
protein and MAPK/ERK signaling pathway-related protein expression.
Data are presented as the mean ± standard deviation (n=3
independent experiments). *P<0.05,
**P<0.01 and ***P<0.001. si-, small
interfering; NC, negative control; t-, total; p-, phosphorylated;
YWHAH, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase
activation protein Eta; ns, not significant (P>0.05).

Figure 9

Western blot assay determines the
expression of autophagy-related proteins and MAPK/ERK signaling
pathway-related proteins in colorectal cancer cells after the
addition of the MAPK/ERK pathway activator U-46619. (A and B)
Western blot assays detected the expression of autophagy-related
proteins and MAPK/ERK signaling pathway-related proteins. (C-F)
Semi-quantitative analysis of the expression levels of
autophagy-related proteins and MAPK/ERK signaling pathway-related
proteins. Data are presented as the mean ± standard deviation (n=3
independent experiments). *P<0.05,
**P<0.01 and ***P<0.001. OE,
overexpression; NC, negative control; t-, total; p-,
phosphorylated; YWHAH, tyrosine 3-monooxygenase/tryptophan
5-monooxygenase activation protein Eta; ns, not significant
(P>0.05).

Figure 10

YWHAH inhibits autophagy through
MAPK/ERK signaling pathway in vivo. (A and B) Western
blotting was used to evaluate the protein expression of ATG7, P62,
LC3II/I, t-ERK1/2, p-ERK1/2 and p-ELK-1 when the nude mice were
injected with CRC cells. (C-F) Semi-quantitative analysis of
autophagy related proteins and MAPK/ERK signaling related proteins
when the nude mice were injected with CRC cells.
*P<0.05, **P<0.01 and
***P<0.001. Data are presented as the mean ± standard
deviation (n=3 independent experiments). YWHAH, tyrosine
3-monooxygenase/tryptophan 5-monooxygenase activation protein Eta;
t-, total; p-, phosphorylated; CRC, colorectal cancer; ATG7,
autophagy related 7; OE, overexpression; si-, small interfering;
NC, negative control; ns, not significant (P>0.05).
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Copy and paste a formatted citation
Spandidos Publications style
Li Q, Wang Y, Yuan Z, Zhai P, Wang J, Zhang C, Shao Z and Xing C: YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion. Int J Mol Med 56: 228, 2025.
APA
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C. ... Xing, C. (2025). YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion. International Journal of Molecular Medicine, 56, 228. https://doi.org/10.3892/ijmm.2025.5669
MLA
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C., Shao, Z., Xing, C."YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion". International Journal of Molecular Medicine 56.6 (2025): 228.
Chicago
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C., Shao, Z., Xing, C."YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion". International Journal of Molecular Medicine 56, no. 6 (2025): 228. https://doi.org/10.3892/ijmm.2025.5669
Copy and paste a formatted citation
x
Spandidos Publications style
Li Q, Wang Y, Yuan Z, Zhai P, Wang J, Zhang C, Shao Z and Xing C: YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion. Int J Mol Med 56: 228, 2025.
APA
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C. ... Xing, C. (2025). YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion. International Journal of Molecular Medicine, 56, 228. https://doi.org/10.3892/ijmm.2025.5669
MLA
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C., Shao, Z., Xing, C."YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion". International Journal of Molecular Medicine 56.6 (2025): 228.
Chicago
Li, Q., Wang, Y., Yuan, Z., Zhai, P., Wang, J., Zhang, C., Shao, Z., Xing, C."YWHAH‑driven autophagy via MAPK/ERK signaling enhances CRC cell migration and invasion". International Journal of Molecular Medicine 56, no. 6 (2025): 228. https://doi.org/10.3892/ijmm.2025.5669
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