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Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4

  • Authors:
    • Chaoyi Xia
    • Jingshu Min
  • View Affiliations / Copyright

    Affiliations: Zhejiang Provincial Engineering Research Center of New Technologies and Applications for Targeted Therapy of Major Diseases, College of Life Sciences and Medicine, Zhejiang Sci‑Tech University, Hangzhou, Zhejiang 310018, P.R. China
    Copyright: © Xia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 551
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    Published online on: September 24, 2025
       https://doi.org/10.3892/ol.2025.15297
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Abstract

The four major metabolic pathways in cells, including amino acid, glucose, nucleotide and lipid metabolism, constitute the fundamental framework of cellular metabolic networks. However, the intricate reciprocal regulatory mechanisms between amino acid metabolism and lipid metabolism remain poorly understood. In the present study, an essential amino acid‑deficient culture medium was established and applied to cell cultures. Subsequent analysis using flow cytometry demonstrated that methionine deprivation significantly reduced cellular lipid accumulation and impaired the fatty acid uptake capacity. Mechanistic investigations using western blotting revealed that the methionine‑methionine adenosyltransferase 2A (MAT2A)‑S‑adenosylmethionine (SAM) metabolic axis regulates the protein expression of ACSL4, a critical enzyme governing fatty acid uptake. Notably, the results indicated that the protein expression of MAT2A may be tightly regulated by SAM through a feedback mechanism, ensuring homeostasis of the methionine‑MAT2A‑SAM metabolic axis. In summary, the findings of the present study demonstrate that the methionine‑MAT2A‑SAM axis modulates lipid metabolism by regulating ACSL4 expression and function.
View Figures

Figure 1

Methionine restriction downregulates
cellular lipid levels. (A) HT1080 cells were cultured without
leucine, isoleucine, lysine, methionine, phenylalanine, tyrosine,
tryptophan, valine, histidine or arginine as indicated for 24 h,
and the intracellular lipids were determined by BODIPY staining
coupled with flow cytometry (n=3). (B) HT1080 cells were cultured
without leucine, isoleucine, lysine, methionine, phenylalanine,
tyrosine, tryptophan, valine, histidine, or arginine as indicated
for 24 h, and fatty acid uptake capacity was determined by
BODIPY-C12 staining coupled with flow cytometry (n=3). (C) Western
blot analysis of ACSL4 expression in HT1080 cells treated with
medium without leucine, isoleucine, lysine, methionine,
phenylalanine, tyrosine, tryptophan, valine, histidine or arginine
as indicated for 24 h. (D) Western blot analysis of ACSL4
expression in MEFs cells treated with medium without methionine for
12 h. Data are presented as the mean ± SD with P-values determined
by one-way ANOVA followed by Tukey's Honestly Significant
Difference. *P<0.05, **P<0.01, ***P<0.001,
****P<0.0001. AA, all amino acids; L, isoleucine; I, isoleucine;
K, lysine; M, methionine; F, phenylalanine; T, threonine; W,
tryptophan; V, valine; H, histidine; R, arginine; ACSL4, acyl-CoA
synthetase long chain family member 4; MEFs, mouse embryonic
fibroblasts; Met, methionine.

Figure 2

Methionine regulates ACSL4 through
SAM. (A) Western blot analysis of ACSL4 expression in HT1080 cells
cultured ± methionine or ± SAM as indicated for 24 h. The
ACSL4 mRNA levels in HT1080 cells cultured (B) ± methionine
and ± SAM (C) or ± ADOX as indicated for 24 h were analyzed using
reverse transcription-quantitative PCR (n=3). (D) Western blot
analysis of ACSL4 expression in H1299 cells cultured ± methionine,
± SAM or ± ADOX as indicated for 48 h. (E) H1299 cells were
cultured ± methionine, ± SAM or ± ADOX as indicated for 48 h, and
fatty acid uptake capacity was determined by BODIPY-C12 staining
coupled with flow cytometry (n=3). Data are presented as mean ± SD
with P-values determined by one-way ANOVA followed by Tukey's
Honestly Significant Difference. *P<0.05, **P<0.01,
****P<0.0001. ACSL4, acyl-CoA synthetase long chain family
member 4; SAM, S-adenosylmethionine; ADOX, adenosine dialdehyde;
Met, methionine.

Figure 3

MAT2A can be dynamically regulated by
the level of SAM. (A) Western blot analysis of MAT2A expression in
HT1080 cells treated with medium without leucine, isoleucine,
lysine, methionine, phenylalanine, tyrosine, tryptophan, valine,
histidine, or arginine as indicated for 24 h. (B) Western blot
analysis of MAT2A expression in DU145 cells treated with medium
without methionine as indicated for 0, 3, 6, 9, 12, 24, 30 and 36
h. (C) Western blot analysis of MAT2A expression in DU145 cells
cultured ± methionine, ± SAM, ± SAH or ± Hcy as indicated for 30 h.
(D) Western blot analysis of MAT2A expression in H1299 cells
cultured ± methionine, ± SAM or ± ADOX, as indicated for 48 h. The
(E) MAT2A and (F) MAT2B mRNA levels in MEFs and
HT1080 and H1299 cells cultured ± methionine, ± SAM or ± ADOX as
indicated for 12 h, were analyzed using reverse
transcription-quantitative PCR (n=3). Data are presented as mean ±
SD with P-values determined by one-way ANOVA followed by Tukey's
Honestly Significant Difference. **P<0.01, ***P<0.001,
****P<0.0001. ns, non-significant; AA, all amino acids; L,
isoleucine; I, isoleucine; K, lysine; M, methionine; F,
phenylalanine; T, threonine; W, tryptophan; V, valine; H,
histidine; R, arginine; MEFs, mouse embryonic fibroblasts; SAM,
S-adenosylmethionine; ADOX, adenosine dialdehyde; SAH,
S-adenosylhomocysteine; Hcy, homocysteine; MAT2A/B, methionine
adenosyltransferase 2A/B; Met, methionine.

Figure 4

Knocking down MAT2A expression
reduces the intracellular lipid levels. (A) HT1080 cells were
cultured ± methionine as indicated for 20 h, and the intracellular
lipids were determined by BODIPY staining coupled with flow
cytometry (n=3). (B) HT1080 cells were cultured ± methionine as
indicated for 12 h, and fatty acid uptake capacity was determined
by BODIPY-C12 staining coupled with flow cytometry (n=3). (C)
Western blot analysis of MAT2A expression in HT1080 cells
transfected with siMAT2A or siNC. (D) The intracellular lipids in
HT1080 cells transfected with siMAT2A or siNC were determined by
BODIPY staining coupled with flow cytometry (n=3). (E) Fatty acid
uptake capacity in HT1080 cells transfected with siMAT2A or siNC
was determined by BODIPY-C12 staining coupled with flow cytometry
(n=3). Data are presented as mean ± SD, with P-values determined by
one-way ANOVA followed by Tukey's Honestly Significant Difference.
*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. MAT2A,
methionine adenosyltransferase 2A; si, small interfering (RNA); NC,
negative control; Met, methionine.

Figure 5

Methionine restriction suppresses
tumor cell proliferation by inhibiting ACLS4. (A) Analysis of
ACSL4 expression in tumor and adjacent tissues based on TCGA
database. (B) Analysis of the correlation between ACSL4 expression
and overall survival based on TCGA database through GEPIA2
(http://gepia2.cancer-pku.cn/). (C)
Western blot analysis of ACSL4 expression in HT1080 cells treated
with medium without methionine as indicated for 12 h. (D) The
number of HT1080 cells after methionine deprivation as indicated
for 36 h were counted using a microscope and cell counter (n=3).
(E) HT1080 cells were cultured ± methionine as indicated for 36 h
and the lipid ROS level was determined by BODIPY-C11 staining
coupled with flow cytometry (n=3). (F) HT1080 cells were cultured ±
methionine as indicated for 36 h and cell death was determined by
PI staining coupled with flow cytometry (n=3). (G) HT1080 cells
were cultured ± methionine as indicated for 36 h and cell apoptosis
was determined by PI staining coupled with flow cytometry (n=3).
Data are presented as mean ± SD, with P-values determined by
one-way ANOVA followed by Tukey's Honestly Significant Difference.
*P<0.05, **P<0.01, ***P<0.001. ns, non-significant; TCGA,
The Cancer Genome Atlas; TPM, transcripts per million; ESCA,
esophageal carcinoma; LIHC, liver hepatocellular carcinoma; PAAD,
pancreatic adenocarcinoma; STAD, stomach adenocarcinoma; T, tumor;
N, normal; HR, hazard ratio; ACSL4, acyl-CoA synthetase long chain
family member 4; SSC, side Scatter; ROS, reactive oxygen species;
PI, propidium iodide; Met, methionine.
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Copy and paste a formatted citation
Spandidos Publications style
Xia C and Min J: Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4. Oncol Lett 30: 551, 2025.
APA
Xia, C., & Min, J. (2025). Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4. Oncology Letters, 30, 551. https://doi.org/10.3892/ol.2025.15297
MLA
Xia, C., Min, J."Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4". Oncology Letters 30.6 (2025): 551.
Chicago
Xia, C., Min, J."Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4". Oncology Letters 30, no. 6 (2025): 551. https://doi.org/10.3892/ol.2025.15297
Copy and paste a formatted citation
x
Spandidos Publications style
Xia C and Min J: Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4. Oncol Lett 30: 551, 2025.
APA
Xia, C., & Min, J. (2025). Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4. Oncology Letters, 30, 551. https://doi.org/10.3892/ol.2025.15297
MLA
Xia, C., Min, J."Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4". Oncology Letters 30.6 (2025): 551.
Chicago
Xia, C., Min, J."Methionine‑MAT2A‑SAM axis controls lipid levels by regulating ACSL4". Oncology Letters 30, no. 6 (2025): 551. https://doi.org/10.3892/ol.2025.15297
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