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Article Open Access

STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma

  • Authors:
    • Haoyang Hu
    • Haozhe Zhang
    • Shuai Han
    • Jianli Chen
    • Ying Xie
  • View Affiliations / Copyright

    Affiliations: Second Department of Hepatobiliary and Pancreatic Surgery, North China University of Science and Technology Affiliated Hospital, Tangshan, Hebei 063000, P.R. China, Department of Urology, North China University of Science and Technology Affiliated Hospital, Tangshan, Hebei 063000, P.R. China, Laboratory Animal Center, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China
    Copyright: © Hu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 38
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    Published online on: December 4, 2025
       https://doi.org/10.3892/ijmm.2025.5709
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Abstract

Hepatocellular carcinoma (HCC) treatment remains challenging due to the prevalence of metastasis and chemotherapy resistance. Mitochondrial stomatin‑like protein 2 (STOML2), which is upregulated in various solid tumors, is associated with a poor prognosis; however, its biological function and molecular mechanism in HCC remain unclear. The present study aimed to elucidate the oncogenic mechanism of STOML2 in HCC and to explore its potential as a therapeutic target. Firstly, STOML2 expression in HCC and matched normal liver tissues was analyzed. In addition, STOML2‑knockdown (HCCLM3‑short hairpin RNA‑STOML2) and ‑overexpression (Huh7‑STOML2) cell models were established. Wound healing, Cell Counting Kit‑8 and Transwell assays, and flow cytometry were performed to assess cell proliferation, invasion, migration and apoptosis in vitro. Furthermore, the biological function of STOML2 was confirmed in vivo. Co‑immunoprecipitation (co‑IP) and immunofluorescence staining were conducted to validate the interaction of STOML2 with prohibitin (PHB) following the prediction of binding partners. Downstream pathways regulated by STOML2 were identified using western blotting and were further investigated using the RAF1 inhibitor sorafenib. The present study revealed that STOML2 expression was significantly upregulated in HCC tissues and metastatic lesions, and was associated with poor patient prognosis. The in vitro experiments showed that STOML2 overexpression promoted proliferation, invasion, migration and autophagy, while inhibiting apoptosis in Huh7 cells. Conversely, STOML2 knockdown reversed these phenotypic changes. Furthermore, co‑IP confirmed the direct interaction between STOML2 and PHB, which activated the RAF/MEK/ERK signaling pathway. The in vivo experiments further confirmed that STOML2 overexpression significantly accelerated tumor growth, whereas STOML2 or PHB knockdown inhibited tumor progression. In addition, sorafenib treatment suppressed STOML2‑mediated cell migration and the expression of autophagy‑related proteins by blocking the MAPK pathway. These findings elucidated the molecular mechanism by which STOML2 promotes the malignant progression of HCC and demonstrated that targeted inhibition of the PHB‑MAPK pathway may reverse the pro‑tumorigenic effects of STOML2. STOML2 may serve as both a prognostic biomarker and a therapeutic target in HCC. The current study provides a theoretical foundation for individualized treatment in patients with HCC and high STOML2 expression.
View Figures

Figure 1

STOML2 expression is upregulated in
HCC tissues. (A) Overall survival analysis using The Cancer Genome
Atlas dataset showed that patients with HCC exhibiting high STOML2
expression had a significantly worse prognosis. P=0.017, log-rank
test. (B and C) STOML2 expression levels across various cancer
types determined using public datasets from the Gene Expression
Omnibus. (B) GSE40367 dataset comparing normal liver tissue,
non-metastatic HCC, HCC with lung metastasis and HCC with adrenal
metastasis (*P<0.05, **P<0.01,
****P<0.0001, one-way ANOVA followed by Tukey's post
hoc test, n=6, 10, 12 and 6, respectively). Data are presented as
median with interquartile range. (C) GSE14520 dataset comparing HCC
tissues and normal liver tissues (****P<0.0001,
unpaired Student's t-test, n=65 and 50, respectively). Data are
presented as median with interquartile range. (D) STOML2 protein
expression in 72 paired HCC tissues (denoted as T) and adjacent
normal tissues (denoted as N) was evaluated using IHC with tissue
microarrays. (E) Semi-quantitative comparison of IHC staining (IHC
score) for tumor tissues vs. adjacent normal tissues (n=72 pairs).
Individual paired data points are shown as scatter plots, with
lines connecting each tumor tissue to its matched adjacent normal
tissue. The bars represent the median IHC score for each group,
with error bars indicating the interquartile range.
***P<0.001, Wilcoxon signed-rank test. STOML2
expression was assessed in eight randomly selected paired HCC and
normal tissues using (F) western blotting and (G) reverse
transcription-quantitative polymerase chain reaction (n=8 pairs).
(F) Representative images from three independent experiments are
shown and (G) data were obtained from three independent
experiments. ****P<0.0001, paired Student's t-test.
Labels S1-S8 represent eight randomly selected paired HCC and
adjacent normal tissue samples from individual patients. HCC,
hepatocellular carcinoma; IHC, immunohistochemistry; STOML2,
stomatin-like protein 2.

Figure 2

STOML2 promotes proliferation,
migration, invasion and autophagy, and inhibits apoptosis in HCC
cells. (A) STOML2 expression levels in five HCC cell lines and
normal hepatocytes were determined using western blotting and
RT-qPCR. Representative western blot images and quantitative
RT-qPCR data (mean ± SD) from three independent experiments are
shown (****P<0.0001, one-way ANOVA followed by
Tukey's post-hoc tests). (B) Knockdown efficiency in HCCLM3 cells
and overexpression efficiency in Huh7 cells were evaluated using
western blotting and RT-qPCR. Representative western blot images
and quantitative RT-qPCR data (mean ± SD) from three independent
experiments are shown (***P<0.001,
**P<0.01, comparisons between two groups were
analyzed by unpaired Student's t-test, whereas comparisons among
three groups were analyzed by one-way ANOVA followed by Tukey's
post-hoc test). (C) Cell Counting Kit-8 assay showing the
proliferation of STOML2-knockdown HCCLM3 cells,
STOML2-overexpressing Huh7 cells and their respective controls.
Data are presented as mean ± SD from three independent experiments
(****P<0.0001, comparisons between two groups were
analyzed by unpaired Student's t-test, whereas comparisons among
three groups were analyzed by one-way ANOVA followed by Tukey's
post-hoc test). (D) Wound healing assay assessing the migratory
ability of STOML2-overexpressing or -knockdown HCC cells.
Representative images from three independent experiments are shown.
(E) Cell cycle distribution analysis of STOML2-knockdown HCCLM3
cells, STOML2-overexpressing Huh7 cells and their respective
controls. Representative images from three independent experiments
are shown. (F) Flow cytometric analysis of apoptosis rates of
STOML2-overexpressing or -knockdown HCC cells along with their
respective controls (***P<0.001, comparisons between
two groups were analyzed by unpaired Student's t-test, whereas
comparisons among three groups were analyzed by one-way ANOVA
followed by Tukey's post-hoc test). Representative flow cytometry
plots and quantitative data (mean ± SD) from three independent
experiments are shown. (G) Transwell assay measuring the invasive
capacity of HCC cells following STOML2 overexpression or knockdown
(***P<0.001, **P<0.01, comparisons
between two groups were analyzed by unpaired Student's t-test,
whereas comparisons among three groups were analyzed by one-way
ANOVA followed by Tukey's post-hoc test). Data are presented as
mean ± SD. (H) Western blotting of autophagy markers p62 and
Beclin1 in STOML2-overexpressing or -knockdown HCC cells.
Representative western blot images from three independent
experiments are shown. HCC, hepatocellular carcinoma; RT-qPCR,
reverse transcription-quantitative polymerase chain reaction; Scr,
scramble; sh, short hairpin; STOML2, stomatin-like protein 2.

Figure 3

STOML2 promotes hepatocellular
carcinoma growth and progression in vivo. (A-E) Subcutaneous
implantation of STOML2-overexpressing Huh7 cells and
STOML2-knockdown HCCLM3 cells along with their control cells in
BALB/c nude mice. (A) Each mouse received 3×106 cells as
a subcutaneous injection at the dorsal region of their neck. Tumors
were harvested 25 days post-injection. (B) Tumor growth curves
obtained at the indicated time points (****P<0.0001,
comparisons between two groups were analyzed by unpaired Student's
t-test, whereas comparisons among three or more groups were
analyzed by two-way ANOVA followed by Tukey's post hoc test). Data
are presented as mean ± SD. (C) Final tumor mass measured upon
removal (****P<0.0001, unpaired Student's t-test or
one-way ANOVA and Tukey's post hoc test for multiple comparisons).
Data are presented as mean ± SD. (D) Representative
immunohistochemistry images showing a positive association between
STOML2 and Ki-67 expression in tumor tissues. Quantitative data are
presented as mean ± SD from multiple fields of view
(****P<0.0001, ***P<0.001, one-way
ANOVA followed by Tukey's post hoc test for multiple comparisons).
(E) Representative hematoxylin and eosin staining of tumor
sections. Images are representative of tumors from each group. (F)
Co-immunoprecipitation assay demonstrating interaction between
STOML2 and PHB in vitro. Representative western blot images
from three independent experiments are shown. (G)
Immunofluorescence staining showing colocalization of STOML2
(green), PHB (red) and DAPI (blue) in HCCLM3, HepG2 and Huh7 cells.
Representative images from three independent experiments are shown.
PHB, prohibitin; Scr, scramble; sh, short hairpin; STOML2,
stomatin-like protein 2.

Figure 4

PHB knockdown inhibits STOML2-induced
proliferation and invasion while promoting pro-apoptotic autophagy
in HCC cells. (A) Western blot analysis of STOML2, PHB and key
proteins of the MAPK signaling pathway in Huh7-STOML2,
HCCLM3-shSTOML2 and their respective control cells. Representative
western blot images from three independent experiments are shown.
(B) Wound healing assay (scale bar, 25 μm), (C) cell cycle
analysis and (D) Transwell assay (****P<0.0001,
unpaired Student's t-test) demonstrating that PHB knockdown
attenuated the cell cycle progression, migration and invasion of
STOML2-overexpressing Huh7 cells. Representative images and
quantitative analysis (mean ± SD) from three independent
experiments are shown. (E) Huh7-STOML2-Scr control and
Huh7-STOML2-shPHB cells were subcutaneously injected into BALB/c
nude mice. Each mouse was injected with 3×106 cells in
the dorsal region of their neck. (F) Flow cytometric analysis
showing that PHB knockdown promoted apoptosis in
STOML2-overexpressing Huh7 cells. Representative flow cytometry
plots and quantitative data (mean ± SD) from three independent
experiments are shown. (unpaired Student's t-test,
***P<0.001). (G) Tumor growth curves obtained at the
indicated time points. Data are presented as mean ± SD
(****P<0.0001, unpaired Student's t-test). (H)
Western blotting of STOML2, PHB and MAPK signaling pathway proteins
in Huh7-STOML2 cells with or without PHB knockdown. Representative
western blot images from three independent experiments are shown.
(I) Tumor mass measured at removal. ****P<0.0001,
unpaired Student's t-test. Data are presented as mean ± SD. (J)
Western blotting of p62 and Beclin1 in Huh7-STOML2 cells with or
without PHB knockdown. Representative western blot images from
three independent experiments are shown. p-, phosphorylated; PHB,
prohibitin; Scr, scramble; sh, short hairpin; STOML2, stomatin-like
protein 2.

Figure 5

Sorafenib (an RAF1 inhibitor)
significantly attenuated STOML2-induced cell cycle progression,
migration and autophagy, while promoting apoptosis in HCC cells.
(A) Cell cycle distribution analysis of Huh7-STOML2 and control
cells following treatment with sorafenib (20 μM, 24 h) or
DMSO. Representative images from three independent experiments are
shown. (B) Wound healing assay showing the migratory capacity of
Huh7-STOML2 cells treated with sorafenib or DMSO. Representative
images from three independent experiments are shown. (C) Flow
cytometric analysis of apoptosis in STOML2-overexpressing Huh7 and
control cells after treatment with sorafenib or DMSO (unpaired
Student's t-test, ****P<0.0001). Representative flow
cytometry plots and quantitative data (mean ± SD) from three
independent experiments are shown. (D) Transwell assay showing the
invasive capacity of Huh7-STOML2 and control cells following
sorafenib treatment or DMSO. Data are presented as mean ± SD
(unpaired Student's t-test, ***P<0.001). Western blot
analysis of (E) MAPK signaling pathway proteins, and (F) p62 and
Beclin1 in Huh7-STOML2 cells after treatment with sorafenib or
DMSO. Representative images from three independent experiments are
shown. p-, phosphorylated; STOML2, stomatin-like protein 2.
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Copy and paste a formatted citation
Spandidos Publications style
Hu H, Zhang H, Han S, Chen J and Xie Y: STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma. Int J Mol Med 57: 38, 2026.
APA
Hu, H., Zhang, H., Han, S., Chen, J., & Xie, Y. (2026). STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma. International Journal of Molecular Medicine, 57, 38. https://doi.org/10.3892/ijmm.2025.5709
MLA
Hu, H., Zhang, H., Han, S., Chen, J., Xie, Y."STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma". International Journal of Molecular Medicine 57.2 (2026): 38.
Chicago
Hu, H., Zhang, H., Han, S., Chen, J., Xie, Y."STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma". International Journal of Molecular Medicine 57, no. 2 (2026): 38. https://doi.org/10.3892/ijmm.2025.5709
Copy and paste a formatted citation
x
Spandidos Publications style
Hu H, Zhang H, Han S, Chen J and Xie Y: STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma. Int J Mol Med 57: 38, 2026.
APA
Hu, H., Zhang, H., Han, S., Chen, J., & Xie, Y. (2026). STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma. International Journal of Molecular Medicine, 57, 38. https://doi.org/10.3892/ijmm.2025.5709
MLA
Hu, H., Zhang, H., Han, S., Chen, J., Xie, Y."STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma". International Journal of Molecular Medicine 57.2 (2026): 38.
Chicago
Hu, H., Zhang, H., Han, S., Chen, J., Xie, Y."STOML2 interacts with PHB to activate the MEK/ERK signaling pathway and mediates autophagy‑related proteins in the progression of hepatocellular carcinoma". International Journal of Molecular Medicine 57, no. 2 (2026): 38. https://doi.org/10.3892/ijmm.2025.5709
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