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Article Open Access

Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH

  • Authors:
    • Xixi Wei
    • Yang Wang
    • Wanlong Zhao
    • Wenqian Yang
    • Jiaping Tang
    • Baosheng Zhao
    • Yuzhen Liu
  • View Affiliations / Copyright

    Affiliations: Henan Key Laboratory of Neurorestoratology, Life Science Research Center, The First Affiliated Hospital of Henan Medical University, Weihui, Henan 453100, P.R. China, Department of Thoracic Surgery, The First Affiliated Hospital of Henan Medical University, Weihui, Henan 453100, P.R. China
    Copyright: © Wei et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 73
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    Published online on: July 25, 2025
       https://doi.org/10.3892/ijo.2025.5779
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Abstract

Glioma is a common and aggressive malignant brain tumor. Despite advances in research, the mechanisms driving glioma initiation and progression remain incompletely understood. The present study aimed to assess the role of acetyl‑CoA carboxylase 1 (ACC1) in glioma, focusing on its mechanistic function in U251 cells and its clinical significance. ACC1 expression was first assessed in four glioma cell lines and then the effects on cellular functions were evaluated. Based on the finding that ACC1 knockdown altered the phenotype of U251 cells, potentially through modulation of succinate dehydrogenase (SDH) activity, further mechanistic assessments were performed. Finally, the association between ACC1 expression and patient prognosis was analyzed. The results demonstrated that ACC1 overexpression inhibited proliferation, migration and invasion in U87 cells. Conversely, ACC1 knockdown promoted these processes in U251, T98G and LN229 cells. Mechanistically, in U251 cells, ACC1 knockdown increased acetyl‑CoA levels, enhancing substrate availability for P300. This led to upregulation of DNA methyltransferase 1 (DNMT1), hypermethylation of the SDH promoter and subsequent SDH downregulation. The resulting increase in reactive oxygen species (ROS) levels promoted U251 cell migration and invasion. Analysis of clinical data revealed a significant correlation between low ACC1 expression and poor survival outcomes in patients with glioma. These findings suggest that ACC1 functions as a tumor suppressor in glioma. Its downregulation promotes a pro‑tumorigenic phenotype via the acetyl‑CoA/P300/DNMT1/SDH/ROS pathway, highlighting its potential as a prognostic marker and therapeutic target. This underscores the importance of developing personalized treatment strategies targeting ACC1 in glioma.
View Figures

Figure 1

Effects of ACC1 modulation on glioma
cell migration/invasion. (A) WB of ACC1 in U87-OE cells. (B) Images
of Transwell migration and invasion assays in U87-OE cells. (C)
Quantification of Transwell assay findings in U87-OE cells. (D)
Wound-healing images of U87-OE cells. (E) Quantification of
wound-healing assay findings in U87-OE cells. (F) WB of ACC1 in
U251-KD cells. (G) Images of Transwell migration and invasion
assays in U251-KD cells. (H) Quantification of Transwell assay
findings in U251-KD cells. (I) Wound-healing images of U251-KD
cells. (J) Quantification of wound-healing assay findings in
U251-KD cells. (K) WB of ACC1 in T98G-KD cells. (L) Images of
Transwell migration and invasion assays in T98G-KD cells. (M)
Quantification of Transwell assay findings in T98G-KD cells. (N)
Wound-healing images of T98G-KD cells. (O) Quantification of
wound-healing assay findings in T98G-KD cells. (P) WB of ACC1 in
LN229-KD cells. (Q) Images of Transwell migration and invasion
assays in LN229-KD cells. (R) Quantification of Transwell assay
findings in LN229-KD cells. (S) Wound-healing images of LN229-KD
cells. (T) Quantification of wound-healing assay findings in
LN229-KD cells. Scale bars, 100 μm. Error bars represent the
mean ± standard deviation from three independent experiments.
*P<0.05; **P<0.01;
***P<0.001. ACC1, acetyl-CoA carboxylase 1; WB,
western blotting; OE, overexpression; KD, knockdown; sh, short
hairpin; NC, negative control.

Figure 2

KD of ACC1 in U251 cells promotes
cell migration and invasion by down-regulating SDH and thus
elevating ROS level. (A) WB of ACC1, SDHA and SDHB after ACC1 KD.
(B) Semi-quantification of ACC1, SDHA and SDHB protein levels after
ACC1 KD. (C) Quantification of SDH activity following ACC1 KD. (D)
Flow cytometric analysis of ROS after ACC1 KD. (E) Quantification
of cellular ROS fluorescence signals after ACC1 KD. (F) Flow
cytometric analysis of ROS after NAC treatment. (G) Quantification
of cellular ROS fluorescence signals after NAC treatment. (H)
Images of Transwell migration and invasion assays after NAC
treatment. (I) Quantification of Transwell migration/invasion assay
findings after NAC treatment. (J) Wound-healing images of cells
after NAC treatment. (K) Quantification of wound-healing assay
after NAC treatment. (L) WB of ACC1, vimentin, fibronectin and
PAI-1 after NAC treatment. (M) Semi-quantification of ACC1,
fibronectin, vimentin and PAI-1 protein levels after NAC treatment.
Scale bars, 100 μm. Error bars represent the mean ± standard
deviation from three independent experiments.
*P<0.05; **P<0.01;
***P<0.001. #P<0.05;
##P<0.01; ###P<0.001. ACC1, acetyl-CoA
carboxylase 1; SDH, succinate dehydrogenase; ROS, reactive oxygen
species; NAC, N-acetyl-cysteine; PAI-1, plasminogen activator
inhibitor-1; sh, short hairpin; NC, negative control; OD, optical
density; WB, western blotting; KD, knockdown.

Figure 3

KD of ACC1 in U251 cells increases
DNMT1 expression, leading to hypermethylation of SDH and subsequent
upregulation of ROS levels. (A) RT-qPCR of DNMT1, DNMT3A and DNMT3B
mRNA levels after ACC1 KD. (B) WB of ACC1 and DNMT1 after ACC1 KD.
(C) Semi-quantification of ACC1 and DNMT1 protein levels after ACC1
KD. (D) RT-qPCR of SDHA, SDHB, SDHC and SDHD mRNA levels after
treatment with Aza. (E) WB of ACC1, SDHA and SDHB after Aza
treatment. (F) Semi-quantification of ACC1, SDHA and SDHB protein
levels after Aza treatment. (G) Methylation-specific PCR of SDHB
promoter methylation after Aza treatment. (H) Semi-quantification
of SDHB promoter methylation levels after Aza treatment. (I) Flow
cytometric analysis of ROS after Aza treatment. (J) Quantification
of cellular ROS fluorescence signals after Aza treatment. (K)
Images of Transwell migration and invasion assays after Aza
treatment. (L) Quantification of Transwell migration/invasion
assays after Aza treatment. (M) Wound-healing images after Aza
treatment. (N) Wound-healing assay quantification after Aza
treatment. (O) WB of ACC1, vimentin, fibronectin and PAI-1 after
Aza treatment. (P) Semi-quantification of ACC1, vimentin,
fibronectin and PAI-1 protein levels after Aza treatment. Scale
bars, 100 μm. Error bars represent the mean ± standard
deviation from three independent experiments.
*P<0.05; **P<0.01;
***P<0.001. #P<0.05;
##P<0.01; ###P<0.001. ACC1, acetyl-CoA
carboxylase 1; DNMT, DNA methyltransferase; SDH, succinate
dehydrogenase; ROS, reactive oxygen species; RT-qPCR, reverse
transcription-quantitative PCR; Aza, azacitidine; PAI-1,
plasminogen activator inhibitor-1; U, unmethylated; M, methylated;
WB, western blotting; KD, knockdown; sh, short hairpin; NC,
negative control.

Figure 4

KD of ACC1 induces histone
acetylation via acetyl-CoA elevation, resulting in DNMT1
upregulation. (A) Fluorometric assay of acetyl-CoA levels after
ACC1 KD. (B) WB of ACC1, H3K9ac and H3 after ACC1 KD. (C)
Semi-quantification of H3K9ac/H3 protein levels after ACC1 KD. (D)
RT-qPCR of DNMT1 mRNA levels after treatment with C646. (E) RT-qPCR
of SDHA, SDHB, SDHC and SDHD mRNA levels after treatment with C646.
(F) WB of ACC1, SDHA, SDHB, H3, DNMT1 and H3K9ac after treatment
with C646. (G) Semi-quantification of ACC1, SDHA, SDHB, DNMT1 and
H3K9ac/H3 protein levels after treatment with C646. (H)
Methylation-specific PCR of SDHB promoter methylation after
treatment with C646. (I) Semi-quantification of SDHB promoter
methylation level after treatment with C646. (J) Flow cytometric
analysis of ROS after treatment with C646. (K) Quantification of
cellular ROS fluorescence signals after treatment with C646. (L)
Images of Transwell migration/invasion assays after treatment with
C646. (M) Quantification of Transwell migration/invasion assays
after treatment with C646. (N) Wound-healing images after treatment
with C646. (O) Wound-healing assay quantification after treatment
with C646. (P) WB of ACC1, vimentin, fibronectin and PAI-1 after
treatment with C646. (Q) Semi-quantification of ACC1, vimentin,
fibronectin and PAI-1 protein levels after treatment with C646.
Scale bars, 100 μm. Error bars represent the mean ± standard
from three independent experiments. *P<0.05;
**P<0.01; ***P<0.001.
#P<0.05; ##P<0.01;
###P<0.001. ACC1, acetyl-CoA carboxylase 1; DNMT, DNA
methyltransferase; H3K9ac, histone H3 acetylation at lysine 9; SDH,
succinate dehydrogenase; PAI-1, plasminogen activator inhibitor-1;
U, unmethylated; M, methylated; WB, western blotting; KD,
knockdown; sh, short hairpin; NC, negative control.

Figure 5

KD of ACC1 in U251 cells elevates
DNMT1 expression via P300. (A) RT-qPCR of P300 mRNA after siP300
transfection in wild-type U251 cells. (B) RT-qPCR of P300 mRNA
after siP300 transfection in ACC1 KD cells. (C) RT-qPCR of DNMT1
mRNA after siP300 transfection. (D) RT-qPCR of SDHA, SDHB, SDHC and
SDHD mRNA after siP300 transfection. (E) Methylation-specific PCR
of SDHB promoter methylation after siP300 transfection. (F)
Semi-quantification of SDHB promoter methylation levels after
siP300 transfection. (G) Flow cytometric analysis of ROS after
siP300 transfection. (H) Quantification of cellular ROS
fluorescence signals after siP300 transfection. (I) Images of
Transwell migration/invasion assays after siP300 transfection. (J)
Quantification of Transwell migration/invasion assays after siP300
transfection. (K) Wound-healing images after siP300 transfection.
(L) Wound-healing assay quantification after siP300 transfection.
(M) WB of ACC1, SDHA, SDHB, H3K9ac, P300, DNMT1, vimentin, H3,
fibronectin and PAI-1 after siP300 transfection. (N)
Semi-quantification of ACC1, SDHA, SDHB, H3K9ac/H3, P300, DNMT1,
vimentin, fibronectin and PAI-1 protein levels after siP300
transfection. Scale bars, 100 μm. Error bars represent the
mean ± standard deviation from three independent experiments.
*P<0.05; **P<0.01;
***P<0.001. #P<0.05;
##P<0.01; ###P<0.001. ACC1, acetyl-CoA
carboxylase 1; DNMT, DNA methyltransferase; SDH, succinate
dehydrogenase; si, small interfering; U, unmethylated; M,
methylated; ROS, reactive oxygen species; H3K9ac, histone H3
acetylation at lysine 9; PAI-1, plasminogen activator inhibitor-1;
WB, western blotting; KD, knockdown; RT-qPCR, reverse
transcription-quantitative PCR; sh, short hairpin; NC, negative
control.

Figure 6

Patients with glioma with low ACC1
expression exhibit poor prognosis. (A) Representative IHC staining
of ACC1 on glioma tissue microarray (n=123). (B) Patients with
grade III-IV glioma demonstrated poor OS. (C) Protein expression of
ACC1 in glioma specimens of different grades. (D) Patients with
glioma with low ACC1 expression exhibited poor OS. (E) Correlation
analysis of survival time and ACC1 expression in glioma samples.
(F) TCGA analysis shows downregulated ACC1 mRNA in glioma tissues.
(G) CPTAC analysis shows downregulated ACC1 protein in glioma
tissues. (H) Analysis of CGGA mRNA_array_301 links low ACC1
expression to poor glioma prognosis. (I) Analysis of CGGA
mRNAseq_325 links low ACC1 expression to poor glioma prognosis. (J)
Analysis of CGGA mRNAseq_693 links low ACC1 expression to poor
glioma prognosis. (K) Patients with glioma with low ACC1 expression
demonstrated a high risk of recurrence. Scale bars, 200 μm.
*P<0.05; ***P<0.001. ACC1, acetyl-CoA
carboxylase 1; IHC, immunohistochemistry; OS, overall survival;
TCGA, The Cancer Genome Atlas; CPTAC, Clinical Proteomic Tumor
Analysis Consortium; CGGA, Chinese Glioma Genome Atlas; GBM,
glioblastoma multiforme.

Figure 7

ACC1 knockdown triggers a cascade of
events in U251 cells, increasing acetyl-CoA levels and enhancing
P300 substrate availability. This prompts an increase in DNMT1
expression, leading to hypermethylation of the SDH promoter,
reduced SDH expression and increased ROS levels. Ultimately, these
molecular alterations promote the migration and invasion of U251
cells. Down-regulation of ACC1 is associated with poor prognosis
and a high-grade of glioma. ACC1, acetyl-CoA carboxylase 1; DNMT,
DNA methyltransferase; SDH, succinate dehydrogenase; ROS, reactive
oxygen species; Ac, acetylation.
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Copy and paste a formatted citation
Spandidos Publications style
Wei X, Wang Y, Zhao W, Yang W, Tang J, Zhao B and Liu Y: Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH. Int J Oncol 67: 73, 2025.
APA
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., & Liu, Y. (2025). Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH. International Journal of Oncology, 67, 73. https://doi.org/10.3892/ijo.2025.5779
MLA
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., Liu, Y."Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH". International Journal of Oncology 67.3 (2025): 73.
Chicago
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., Liu, Y."Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH". International Journal of Oncology 67, no. 3 (2025): 73. https://doi.org/10.3892/ijo.2025.5779
Copy and paste a formatted citation
x
Spandidos Publications style
Wei X, Wang Y, Zhao W, Yang W, Tang J, Zhao B and Liu Y: Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH. Int J Oncol 67: 73, 2025.
APA
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., & Liu, Y. (2025). Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH. International Journal of Oncology, 67, 73. https://doi.org/10.3892/ijo.2025.5779
MLA
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., Liu, Y."Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH". International Journal of Oncology 67.3 (2025): 73.
Chicago
Wei, X., Wang, Y., Zhao, W., Yang, W., Tang, J., Zhao, B., Liu, Y."Knockdown of ACC1 promotes migration and invasion of U251 glioma cells by epigenetically suppressing SDH". International Journal of Oncology 67, no. 3 (2025): 73. https://doi.org/10.3892/ijo.2025.5779
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