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Article

MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis

  • Authors:
    • Sibei Guo
    • Yongya Zhang
    • Xue Li
    • Xueru Zhao
    • Yingjuan Liang
    • Xiaohua Li
  • View Affiliations / Copyright

    Affiliations: Department of Ophthalmology, Xinxiang Medical University, Xinxiang, Henan 453003, P.R. China, Henan Provincial People's Hospital, Henan Eye Hospital, Henan Eye Institute, Henan Key Laboratory of Ophthalmology and Visual Science, Zhengzhou, Henan 450003, P.R. China
  • Article Number: 59
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    Published online on: December 4, 2025
       https://doi.org/10.3892/mmr.2025.13769
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Abstract

Proliferative vitreoretinopathy (PVR), a leading complication of retinal detachment with high recurrence rates and no effective pharmacological treatments, is driven by retinal pigment epithelium (RPE) cells through epithelial‑mesenchymal transition (EMT), a process promoted by methyl‑CpG binding protein 2 (MeCP2). There is bidirectional crosstalk between ferroptosis, an iron‑dependent cell death pathway characterized by lipid peroxidation and EMT, suggesting their interaction may influence PVR pathogenesis. However, the mechanistic involvement of ferroptosis in PVR and its interaction with the MeCP2/EMT axis remain poorly understood. In the present study, a scratch assay demonstrated that MeCP2 enhanced ARPE‑19 cell migration, which was markedly suppressed by erastin. Cell Counting Kit‑8 assays and western blot analysis confirmed that Erastin inhibited cell proliferation without triggering apoptosis. Western blotting and corresponding assay kits both revealed that MeCP2 upregulated glutathione peroxidase 4 (GPX4), glutamate‑cysteine ligase modifier subunit and solute carrier family 7 member 11, increased glutathione levels and decreased malondialdehyde and Fe2+ concentrations, indicating ferroptosis suppression. Erastin reversed EMT by reducing fibronectin (FN) and α‑smooth muscle actin (α‑SMA) expression and restoring E‑cadherin, as shown by western blotting. Further investigation revealed that GPX4 activation exacerbated EMT marker expression (FN, α‑SMA and N‑cadherin), while GPX4 inhibition mitigated these effects, confirming that MeCP2 regulates EMT through GPX4‑dependent ferroptosis. Erastin inhibited MeCP2‑driven ARPE‑19 proliferation, migration and EMT via ferroptosis induction, independent of apoptosis. MeCP2 suppressed ferroptosis through GPX4 upregulation, using this pathway to orchestrate EMT, thus revealing a critical GPX4‑dependent mechanism that links ferroptosis to RPE plasticity in PVR. These findings highlighted ferroptosis modulation as a promising therapeutic strategy for PVR.
View Figures

Figure 1

Erastin inhibits ARPE-19 cell
proliferation. The CCK-8 assay demonstrated the effect of Erastin
on cell proliferation. ARPE-19 cells were treated with varying
concentrations of Erastin (2, 4, 8, 10 and 20 µM) for 24 h.
***P<0.001 (P=0.002, 4 µM vs. control), ****P<0.0001 (4, 8,
10, 20 µM vs. control). Data represent four independent experiments
(n=4).

Figure 2

Erastin modulates ARPE-19 cell
migration. Cell migration was quantified using the wound healing
assay. (A) Representative wound healing assays showing (B)
migration of ARPE-19 cells treated with MeCP2 (100 ng/ml, 24 h),
Erastin (10 µM, 24 h), or a combination of MeCP2 (100 ng/ml, 24 h)
and Erastin (10 µM, 24 h). Magnification, ×40. **P<0.01
(P=0.0012, MeCP2 vs. control; P=0.0015, Erastin vs. control;
P=0.0023, Erastin vs. MeCP2 + Erastin), ***P<0.001 (P=0.0008,
MeCP2 vs. MeCP2 + Erastin), ****P<0.0001 (MeCP2 vs. Erastin).
Data represent three independent experiments (n=3). MeCP2,
methyl-CpG binding protein 2.

Figure 3

The effect of MeCP2 on ferroptosis in
ARPE-19 cells. Cells were treated with MeCP2 (100 ng/ml, 24 h),
Erastin (10 µM, 24 h) and a combination of MeCP2 (100 ng/ml, 24 h)
and Erastin (10 µM, 24 h). (A) GSH levels in ARPE-19 cells were
quantified using a GSH assay kit, ***P<0.001 (P=0.0002, MeCP2
vs. control; P=0.0001, MeCP2 + Erastin vs. control),
****P<0.0001 (Erastin vs. control, MeCP2 vs. Erastin, MeCP2 vs.
MeCP2 + Erastin). (B) MDA content was measured using an MDA assay
kit, *P<0.05 (P=0.0138, Erastin vs. control; P=0.0474, Erastin
vs. MeCP2 + Erastin), **P<0.01 (P=0.0037, MeCP2 vs. Erastin).
(C) Intracellular Fe2+ concentrations were determined
using an iron assay kit, *P<0.05(P=0.035, MeCP2 vs. control;
P=0.031, Erastin vs. MeCP2 + Erastin), **P<0.01 (P=0.008,
Erastin vs. control; P=0.009, MeCP2 vs. MeCP2 + Erastin),
***P<0.001 (P=0.0002, MeCP2 vs. Erastin). (D and E) Expression
patterns of ferroptosis-associated proteins GPX4, GCLM and SLC7A11
were analyzed by western blotting. Blots were probed with anti-GCLM
(1:4,000), anti-GPX4 (1:2000), anti-SLC7A11 (1:1,000) and
anti-GAPDH (1:10,000), with GAPDH as the loading control.
Statistical significance compared with the control group:
*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001(GPX4:
P=0.0034, MeCP2 vs. control; P=0.0243, Erastin vs. control;
P<0.0001, MeCP2 vs. Erastin; P=0.0006, MeCP2 vs. MeCP2 +
Erastin. GCLM: P=0.0027, MeCP2 vs. control; P=0.225, Erastin vs.
control; P<0.0001, MeCP2 vs. Erastin; P=0.0011, MeCP2 vs. MeCP2
+ Erastin. SLC7A11: P=0.0323, MeCP2 vs. control; P=0.045, Erastin
vs. control; P=0.0006, MeCP2 vs. Erastin; P=0.0015, MeCP2 vs. MeCP2
+ Erastin). Data represent three independent experiments (n=3).
MeCP2, methyl-CpG binding protein 2; GSH, glutathione; MDA,
malondialdehyde; GPX4, glutathione peroxidase 4; GCLM,
glutamate-cysteine ligase modifier subunit; SLC7A11, solute carrier
family 7 member 11.

Figure 4

Effect of ferroptosis on
MeCP2-Regulated EMT in ARPE-19 Cells. Cells were treated separately
with MeCP2 (100 ng/ml, 24 h), Erastin (10 µM, 24 h) and a
combination of MeCP2 (100 ng/ml, 24 h) and Erastin (10 µM, 24 h).
(A and B) Western blot analysis was performed to examine the
expression patterns of EMT-associated proteins, including FN, α-SMA
and E-cadherin. Blots were probed with anti-FN (1:3,000),
anti-α-SMA (1:1,000), anti-E-cadherin (1:3,000) and anti-GAPDH
(1:10,000), with GAPDH as the loading control. Statistical
significance compared with the control group: *P<0.05,
**P<0.01, ***P<0.001, ****P<0.0001 (FN: P=0.0056, MeCP2
vs. control; P=0.0002, Erastin vs. control; P=0.0087, MeCP2 +
Erastin vs. control; P<0.0001, MeCP2 vs. Erastin, Erastin vs.
MeCP2 + Erastin. E-Ca: P=0.0287, MeCP2 vs. control; P=0.0002,
Erastin vs. control; P=0.0259, MeCP2 + Erastin vs. control;
P<0.0001, MeCP2 vs. Erastin; P=0.0069, MeCP2 vs. MeCP2 +
Erastin; P=0.0092, Erastin vs. MeCP2 + Erastin. α-SMA: P=0.0154,
MeCP2 vs. control; P=0.0414, Erastin vs. control; P=0.0004, MeCP2
vs. Erastin; P=0.0271, MeCP2 vs. MeCP2 + Erastin; P=0.0233, Erastin
vs. MeCP2 + Erastin). n=3. MeCP2, methyl-CpG binding protein 2;
EMT, epithelial-mesenchymal transition; FN, fibronectin; α-SMA,
α-smooth muscle actin; E-Ca, E-cadherin.

Figure 5

The effect of Erastin on apoptosis in
ARPE-19 Cells. Cells were treated with Erastin (10 µM, 24 h). (A
and B) Western blot analysis was performed to examine the
expression patterns of apoptosis-related proteins, including Bax,
Caspase-9 and Cleaved Caspase-9 in ARPE-19 cells. Blots were probed
with anti-Bax (1:1,000), anti-Caspase-9 (1:1,000), anti-Cleaved
Caspase-9 (1:500) and anti-GAPDH (1:10,000), with GAPDH as the
loading control. No statistical significance was observed compared
with the control group. Data are representative of three
independent experiments (n=3). ns, no significance; E-Ca,
E-cadherin.

Figure 6

MeCP2 modulates ferroptosis-mediated
EMT in ARPE-19 cells through GPX4 Regulation. Cells were treated
separately with MeCP2 (100 ng/ml, 24 h), a combination of MeCP2
(100 ng/ml, 24 h) and RSL3 (0.5 µM, 24 h) and a combination of
MeCP2 (100 ng/ml, 24 h) and PKUMDL-LC-101-D04 (65 µM, 24 h). (A)
Representative western blot of (B) EMT-associated proteins (FN,
α-SMA, and N-cadherin) in ARPE-19 cells following GPX4 activation
and inhibition. *P<0.05, **P<0.01, ****P<0.0001 vs.
control. (N-Ca: P=0.0029, MeCP2 vs. MeCP2 + RSL3; P=0.0018, MeCP2
vs. MeCP2 + PKUMDL-LC-101-D04; P<0.0001, MeCP2 + RSL3 vs. MeCP2
+ PKUMDL-LC-101-D04. α-SMA: P=0.0482, MeCP2 vs. MeCP2 + RSL3;
P=0.0219, MeCP2 vs. MeCP2 + PKUMDL-LC-101-D04; P=0.0012, MeCP2 +
RSL3 vs. MeCP2 + PKUMDL-LC-101-D04. FN: P=0.0056, MeCP2 vs. MeCP2 +
RSL3; P=0.0448, MeCP2 vs. MeCP2 + PKUMDL-LC-101-D04; P=0.0221,
MeCP2 + RSL3 vs. MeCP2 + PKUMDL-LC-101-D04). n=3. MeCP2, methyl-CpG
binding protein 2; EMT, epithelial-mesenchymal transition; GPX4,
glutathione peroxidase 4; FN, fibronectin; α-SMA, α-smooth muscle
actin; N-Ca, N-cadherin.
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Copy and paste a formatted citation
Spandidos Publications style
Guo S, Zhang Y, Li X, Zhao X, Liang Y and Li X: MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis. Mol Med Rep 33: 59, 2026.
APA
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., & Li, X. (2026). MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis. Molecular Medicine Reports, 33, 59. https://doi.org/10.3892/mmr.2025.13769
MLA
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., Li, X."MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis". Molecular Medicine Reports 33.2 (2026): 59.
Chicago
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., Li, X."MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis". Molecular Medicine Reports 33, no. 2 (2026): 59. https://doi.org/10.3892/mmr.2025.13769
Copy and paste a formatted citation
x
Spandidos Publications style
Guo S, Zhang Y, Li X, Zhao X, Liang Y and Li X: MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis. Mol Med Rep 33: 59, 2026.
APA
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., & Li, X. (2026). MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis. Molecular Medicine Reports, 33, 59. https://doi.org/10.3892/mmr.2025.13769
MLA
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., Li, X."MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis". Molecular Medicine Reports 33.2 (2026): 59.
Chicago
Guo, S., Zhang, Y., Li, X., Zhao, X., Liang, Y., Li, X."MeCP2 suppresses ferroptosis to drive EMT in retinal pigment epithelial cells: Implications for PVR pathogenesis". Molecular Medicine Reports 33, no. 2 (2026): 59. https://doi.org/10.3892/mmr.2025.13769
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