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HMGB1 mediates low-dose ionizing radiation-induced Wnt/β-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis

  • Authors:
    • Ping Wang
    • Chunnan Piao
    • Dong Yan
    • Yingxue He
    • Yaru Li
    • Li Fan
    • Mei Tian
  • View Affiliations / Copyright

    Affiliations: Department of Nuclear Medicine, PLA 960th Hospital, Jinan, Shandong 250031, P.R. China, China CDC Key Laboratory of Radiation Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, P.R. China
    Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 37
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    Published online on: December 4, 2025
       https://doi.org/10.3892/ijmm.2025.5708
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Abstract

Emerging evidence from our prior investigations has elucidated the dose-dependent regulatory effects of low-dose ionizing radiation on cellular behaviors including proliferation, migration and differentiation in HLE-B3 lens epithelial cells, with concomitant activation of the canonical Wnt/β-catenin signaling cascade. To extend these findings to alternative cellular models, the present study systematically evaluated the biological responses of the well-characterized human lens epithelial cell line SRA01/04 to low-dose ionizing radiation exposure (0.05-0.2 Gy) versus high-dose radiation (0.5-2 Gy), with particular emphasis on temporal dynamics during acute (0-72 h) and chronic (7 days) phases. Mechanistically, lentivirus-mediated RNA interference was employed to establish stable High mobility group box protein 1 (HMGB1)-knockdown cell models, enabling rigorous interrogation of β-catenin subcellular localization and functional readouts under 0, 0.1 and 0.2 Gy γ-ray exposures. Key findings revealed the following: i) low-dose ionizing radiation within the 0.05-0.2 Gy range significantly potentiated SRA01/04 cell proliferation and migration capacity (P<0.05), concomitant with nuclear accumulation of β-catenin; ii) genetic ablation of HMGB1 abolished radiation-induced β-catenin nuclear translocation, resulting in 77% reduction in proliferation rate and 82% suppression of migratory activity compared with wild-type counterparts under equivalent radiation. The experimental evidence identifies HMGB1-mediated signaling as the critical molecular nexus connecting low-dose ionizing radiation exposure to dysregulated Wnt/β-catenin activity in lens epithelium, offering a new therapeutic target for preventing radiation-related cataracts.
View Figures

Figure 1

Effects of different doses of γ-rays
on the proliferation of SRA01/04 cells. (A) Changes in cell
viability at 8-72 h and 7 days after irradiation (n=6). (B) Changes
in cell proliferation ability 0-72 h after irradiation (n=6). Data
represent the mean±SD of 3 independent experiments.
*P<0.05, **P<0.01 and
***P<0.001, compared with the non-irradiated group;
#P<0.05 compared with 2 Gy group, One-way ANOVA. OD,
optical density.

Figure 2

Gap closure assay was used to
investigate migration behavior of SRA01/04 cells after exposure to
different doses of γ-rays. (A) Representative images of gap closure
assay at 0-72 h after scratching. (B-D) Cell gap closure rate at 24
h (B), 48 h (C) and 72 h (D) after irradiation. Magnification,
×200. Data represent the mean ± SD of 3 independent experiments.
*P<0.05 and **P<0.01, compared with the
non-irradiated group, One-way ANOVA.

Figure 3

Transwell assay was used to detect
the effect of different doses of γ-rays on the migration of
SRA01/04 cells. (A-C) Representative images and statistics of
Giemsa staining of migratory cells at 8 h (A), 48 h (B) and 7 days
(C) after irradiation (n=3). Clone formation experiment was used to
detect the proliferation of migratory cells. (D) SRA01/04 cells
migrated to culture dishes 7 days after γ-ray irradiation. (E)
Representative images and quantitative clonogenic analysis of plate
colony formation in SRA01/04 cells (n=3). Scale bar, 50 μm
(×200); Scale bar, 100 μm (×100). Data represent the mean ±
SD of 3 independent experiments. *P<0.05,
**P<0.01 and ***P<0.001, compared with
the non-irradiated group, One-way ANOVA.

Figure 4

Giemsa staining and statistical
images of nucleated cells in the posterior lens capsule of mice at
6 months after irradiation with different doses of γ-rays (n=8).
Scale bar, 250 μm (×40); Scale bar, 25 μm (×400).
Data represent the mean±SD of 8 mice.

Figure 5

Effect of different doses of γ-rays
on the protein expression levels of β-catenin, cyclin D1 and c-Myc
in SRA01/04 cells. (A-C) The protein levels of β-catenin, cyclin D1
and c-Myc were analyzed by western blot at 8 (A), 24 (B) and 48 h
(C) after irradiation. (D-F) The relative protein expression levels
of β-catenin, cyclin D1 and c-Myc were quantitatively determined by
the density method at 8 (D), 24 (E) and 48 (F) after irradiation.
GAPDH was used as a loading control. Data represent the mean ± SD
of 3 independent experiments. *P<0.05,
**P<0.01 and ***P<0.001, compared with
the non-irradiated group, one-way ANOVA.

Figure 6

Immunofluorescence microscopy image
showing the immunolocalization of β-catenin protein in SRA01/04
cells at 24 (A) and 48 h (B) after irradiation with different doses
of γ-rays. This experiment was repeated three times independently.
Scale bar, 20 μm.

Figure 7

Expression levels and quantitative
analysis of HMGB1 in SRA01/04 cells at 24 (A) and 48 h (B) after
irradiation with different doses of γ-rays. (C) Identification of
lentiviral infected stable HMGB1 knockdown cell lines. GAPDH was
used as a loading control. Data represent the mean ± SD of 3
independent experiments. ***P<0.001 compared with the
non-irradiated group, One-way ANOVA. HMGB1, High mobility group box
protein 1.

Figure 8

Effect of HMGB1 on the expression of
β-catenin in SRA01/04 cells after low-dose γ-ray irradiation.
Expression and quantification of total and p-β-catenin in cells at
48 h after irradiation with different doses of γ-rays. GAPDH was
used as a loading control. Data represent the mean±SD of 3
independent experiments. **P<0.01, compared with the
NC group at the same irradiation dose; ##P<0.01 and
###P<0.001, compared with the WT group at the same
irradiation dose, One-way ANOVA. HMGB1, High mobility group box
protein 1. p-β-catenin, phosphorylated β-catenin; WT, wild-type;
NC, negative control; sh-, short hairpin.

Figure 9

Effect of HMGB1 on the localization
of β-catenin in SRA01/04 cells after low-dose γ-ray irradiation.
(A) Non-irradiated group. (B) 0.1 Gy irradiation. (C) 0.2 Gy
irradiation. This experiment was repeated three times
independently. Scale bar, 20 μm. HMGB1, High mobility group
box protein 1; WT, wild-type; NC, negative control; sh-, short
hairpin.

Figure 10

Effects of HMGB1 on the proliferation
and migration of SRA01/04 cells after low-dose γ-ray irradiation.
(A) Effect of HMGB1 on cell viability (n=6). (B) Effect of HMGB1 on
cell proliferative activity (n=6). (C) Representative Giemsa
staining and number statistics of cells migrating to the bottom of
the chamber at 48 h after 0.1 and 0.2 Gy γ-ray irradiation (n=3).
Data represent the mean±SD of 3 independent experiments, Scale bar,
50 μm. *P<0.05, **P<0.01 and
***P<0.001 compared with the NC group at the same
irradiation dose; #P<0.05 and ##P<0.01
compared with the WT group at the same irradiation dose, One-way
ANOVA. HMGB1, High mobility group box protein 1; OD, optical
density; WT, wild-type; NC, negative control; sh-, short
hairpin.
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Copy and paste a formatted citation
Spandidos Publications style
Wang P, Piao C, Yan D, He Y, Li Y, Fan L and Tian M: HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis. Int J Mol Med 57: 37, 2026.
APA
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., & Tian, M. (2026). HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis. International Journal of Molecular Medicine, 57, 37. https://doi.org/10.3892/ijmm.2025.5708
MLA
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., Tian, M."HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis". International Journal of Molecular Medicine 57.2 (2026): 37.
Chicago
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., Tian, M."HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis". International Journal of Molecular Medicine 57, no. 2 (2026): 37. https://doi.org/10.3892/ijmm.2025.5708
Copy and paste a formatted citation
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Spandidos Publications style
Wang P, Piao C, Yan D, He Y, Li Y, Fan L and Tian M: HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis. Int J Mol Med 57: 37, 2026.
APA
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., & Tian, M. (2026). HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis. International Journal of Molecular Medicine, 57, 37. https://doi.org/10.3892/ijmm.2025.5708
MLA
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., Tian, M."HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis". International Journal of Molecular Medicine 57.2 (2026): 37.
Chicago
Wang, P., Piao, C., Yan, D., He, Y., Li, Y., Fan, L., Tian, M."HMGB1 mediates low-dose ionizing radiation-induced Wnt/&beta;-catenin activation in SRA01/04 cells: Mechanistic clues to early cataractogenesis". International Journal of Molecular Medicine 57, no. 2 (2026): 37. https://doi.org/10.3892/ijmm.2025.5708
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