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SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption

  • Authors:
    • Aoxiang Li
    • Xiaolong Li
    • Tuo Wang
    • Jinning Song
  • View Affiliations / Copyright

    Affiliations: Department of Neurosurgery, Xi'an International Medical Center Hospital, Xi'an, Shaanxi 710300, P.R. China, Department of Interventional Radiology, Affiliated Hospital of Yan'an University, Yan'an, Shaanxi 716000, P.R. China, Department of Neurosurgery, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 154
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    Published online on: July 23, 2025
       https://doi.org/10.3892/ijmm.2025.5595
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Abstract

Stromal cell‑derived factor 2 (SDF2) is an endoplasmic reticulum chaperone protein crucial for protein folding. Its role in gliomas is poorly understood. The present study investigated SDF2 expression and function in glioma progression. Our data revealed that the expression of SDF2 was upregulated in glioma tissues. In glioma cell lines, SDF2 promoted cell proliferation and migration, whereas the knockdown of SDF2 (Ad‑shSDF2) induced cell death. Further investigations revealed that the copper chelator tetrathiomolybdate (TTM) could reverse the reduction in cell viability caused by Ad‑shSDF2. Upon SDF2 knockdown, the expression of ATP7A and ATP7B was decreased in glioma cells, whereas the expression of glucose‑regulated protein 78 (GRP78) was increased. Moreover, the proteasome inhibitor MG132 and the silencing of GRP78 effectively blocked the Ad‑shSDF2‑mediated decrease in ATP7A and ATP7B expression, as well as the accumulation of dihydrolipoamide S‑acetyltransferase in mitochondria. In vivo, SDF2 promoted subcutaneous tumor growth in nude mice, an effect that could be reversed by overexpression of GRP78. This reversal was accompanied by an increase in the intra‑tumoral copper ion concentration. In gliomas, SDF2 promotes tumor growth by inhibiting the GRP78‑mediated endoplasmic reticulum‑associated degradation pathway, thereby increasing the expression of ATP7A and ATP7B. This results in reduced intracellular accumulation of copper ions, facilitating tumor progression.
View Figures

Figure 1

Schematic molecular mechanism of SDF2
in ER stress. SDF2, stromal cell-derived factor 2; ER, endoplasmic
reticulum; ERAD, ER-associated degradation; GRP78, glucose-related
protein 78; UPR, unfolded protein response; XBP1, X-box binding
protein 1; ERdj3, endoplasmic reticulum DnaJ heat shock protein
3.

Figure 2

Schematic molecular mechanism of
cuproptosis. GSH, reduced glutathione.

Figure 3

Expression of SDF2 is increased in
glioma tissues. (A and B) Bioinformatic analysis of SDF2 mRNA
levels in glioma tissues compared with those in control tissues
using the GEPIA database. (C) The mRNA level of SDF2 was elevated
in glioma tissues compared with adjacent normal tissues (n=10 for
each group). (D) Immunofluorescent staining revealed that the
intensity of SDF2 was increased in glioma and control tissues (n=3
for each group; scale bar, 50 μm). (E) Western blot analysis
demonstrated that the protein levels of SDF2 were greater in glioma
tissues than in adjacent nontumor tissues (n=5 for each group).

Figure 4

SDF2 promotes malignant proliferation
and migration in glioma cells. (A-C) Reverse
transcription-quantitative PCR analysis revealed that, compared
with those in control cells, SDF2 mRNA levels were significantly
elevated in U87, U251 and HMC3 cells following transfection with
Ad-SDF2 (n=5). (D-F) SDF2 expression was upregulated in U87, U251
and HMC3 cells following transfection with Ad-SDF2 compared with
that in control cells (n=3). (G-I) Cell viability assays
demonstrated that SDF2 overexpression increased the viability of
U87 and U251 cells in a time-dependent manner but did not affect
the viability of HMC3 cells (n=5). (J-M) EdU staining revealed that
the proliferative capacity of U87 and U251 cells was increased
following SDF2 overexpression compared with that of control cells
(n=3; scale bar, 100 μm). (N and O) Transwell migration
assays revealed that the migratory ability of U87 and U251 cells
overexpressing SDF2 was increased (n=3). SDF2, stromal cell-derived
factor 2; NC, negative control.

Figure 5

SDF2 knockdown increases ROS levels
and apoptosis in glioma cells. (A and B) The mRNA levels of SDF2
were lower in U87 and U251 cells transfected with Ad-shSDF2 than in
those transfected with Ad-NC (n=5). (C and D) Ad-shSDF2 decreased
the expression of SDF2 in U87 and U251 cells (n=3). (E and F) Flow
cytometric analysis revealed that the proportions of late apoptotic
and necrotic cells were greater in U87 and U251 cells transfected
with Ad-shSDF2 than in control cells (n=3). (G and H) Intracellular
ROS levels were greater in U87 and U251 cells transfected with
Ad-sh-SDF2 than in those transfected with Ad-NC (n=5). (I and J)
Cell viability assays revealed that only the cuproptosis inhibitor
TTM reversed the decrease in viability induced by Ad-shSDF2 in U87
and U251 cells (n=5). SDF2, stromal cell-derived factor 2; sh-,
short hairpin; TTM, tetrathiomolybdate; NC, negative control.

Figure 6

Knockdown of SDF2 increases
intracellular copper ion accumulation and oxidative stress in
glioma cells. (A) Fluorescence staining suggested that
intracellular copper ion levels were elevated in U87 and U251 cells
following SDF2 knockdown compared with those in control cells (n=3,
scale bar, 50 μm). (B and C) Quantitative assessment of
intracellular copper ion levels revealed significant copper
accumulation in U87 and U251 cells upon SDF2 knockdown (n=5). (D
and E) Western blot analysis revealed that ATP7A, ATP7B, FDX and
lipid-DLAT expression was reduced, whereas DLAT protein expression
did not significantly change (n=3). (F) A decrease in GSH was
identified following SDF2 knockdown in U87 and U251 cells (n=5). (G
and H) Increased ROS and MDA levels were detected in U87 and U251
cells upon SDF2 knockdown (n=3 for each group). SDF2, stromal
cell-derived factor 2; DLAT, dihydrolipoamide S-acetyltransferase;
GSH, reduced glutathione; ROS, reactive oxygen species; MDA,
malondialdehyde; sh-, short hairpin; NC, negative control.

Figure 7

SDF2 knockdown mediates the
downregulation of ATP7A and ATP7B via the endoplasmic
reticulum-associated degradation pathway. (A and C) Reverse
transcription-quantitative PCR analysis revealed that GRP78 mRNA
levels and the XBP1s/XBP1u ratio were increased in U87 and U251
cells following Ad-shSDF2 transfection (n=5). (B and D) Western
blot analysis demonstrated that the protein expression of GRP78,
SDF2L and ERdj3 was increased in U87 and U251 cells after SDF2
knockdown (n=3). (E and F) Western blot analysis revealed that in
U87 and U251 cells, SDF2 knockdown reduced ATP7A and ATP7B
expression compared with that in control cells, whereas
pretreatment with MG132 significantly reversed the
Ad-shSDF2-induced reduction in ATP7A and ATP7B levels (n=3). (G and
H) Immunofluorescent analysis revealed that DLAT aggregation was
increased following Ad-shSDF2 transfection in U87 and U251 cells
(n=3; scale bar, 10 μm). SDF2, stromal cell-derived factor
2; GRP78, glucose-related protein 78; XBP1, X-box binding protein
1; sh-, short hairpin; DLAT, dihydrolipoamide S-acetyltransferase;
ERdj3, endoplasmic reticulum DnaJ heat shock protein 3; NC,
negative control.

Figure 8

GRP78 mediates SDF2-induced
downregulation of ATP7A and ATP7B in glioma cells. (A and D)
Reverse transcription-quantitative PCR analysis showing that,
compared with the control, transfection with si-GRP78 significantly
reduced GRP78 mRNA levels in U87 and U251 cells (n=5). (B and E)
Western blot analysis confirmed that GRP78 knockdown blocked the
Ad-shSDF2-induced downregulation of ATP7A and ATP7B protein levels
in both cell lines (n=3). (C and F) Co-immunoprecipitation analysis
demonstrated that the association between GRP78 and ATP7A/ATP7B was
reduced upon GRP78 knockdown in U87 and U251 cells (n=3). (G and H)
GRP78 knockdown partially reversed the increase in Cu2+
levels and mito-ROS induced by Ad-shSDF2 in U87 and U251 cells
(n=3; scale bar, 100 μm). GRP78, glucose-related protein 78;
SDF2, stromal cell-derived factor 2; si-, small interfering; sh-,
short hairpin; NC, negative control.

Figure 9

Ad-GRP78 reverses Ad-SDF2-induced
tumor growth and metabolic changes in a nude mouse model. (A-C)
Tumor size, volume and weight in nude mice injected with Ad-SDF2 or
Ad-NC (n=5). (D) Immunohistochemical staining demonstrated that
Ad-GRP78 co-expression reversed the Ad-SDF2-induced increase in
Ki-67 positivity (n=5; scale bar, 20 μm). (E) Ad-SDF2
increased GSH levels, whereas Ad-GRP78 restored GSH levels in tumor
tissues (n=5). (F) Ad-SDF2 reduced MDA levels, while Ad-GRP78
partially restored them (n=5). (G and H) Ad-SDF2 increased ATP and
decreased Cu2+ levels, which were restored by Ad-GRP78
co-expression (n=5). (I) Western blot analysis showing that
co-expressing Ad-GRP78 counteracted the Ad-SDF2-induced increases
in ATP7A and ATP7B levels in tumor tissues (n=5). GRP78,
glucose-related protein 78; SDF2, stromal cell-derived factor 2;
GSH, reduced glutathione; MDA, malondialdehyde; NC, negative
control.

Figure 10

Schematic representation of the
mechanism by which SDF2 contributes to glioma development. SDF2,
stromal cell-derived factor 2; GRP78, glucose-related protein 78;
DLAT, dihydrolipoamide S-acetyltransferase; ERdj3, endoplasmic
reticulum DnaJ heat shock protein 3; ERAD, endoplasmic
reticulum-associated degradation; ROS, reactive oxygen species;
URP, unfolded protein response.
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Copy and paste a formatted citation
Spandidos Publications style
Li A, Li X, Wang T and Song J: SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption. Int J Mol Med 56: 154, 2025.
APA
Li, A., Li, X., Wang, T., & Song, J. (2025). SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption. International Journal of Molecular Medicine, 56, 154. https://doi.org/10.3892/ijmm.2025.5595
MLA
Li, A., Li, X., Wang, T., Song, J."SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption". International Journal of Molecular Medicine 56.4 (2025): 154.
Chicago
Li, A., Li, X., Wang, T., Song, J."SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption". International Journal of Molecular Medicine 56, no. 4 (2025): 154. https://doi.org/10.3892/ijmm.2025.5595
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Spandidos Publications style
Li A, Li X, Wang T and Song J: SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption. Int J Mol Med 56: 154, 2025.
APA
Li, A., Li, X., Wang, T., & Song, J. (2025). SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption. International Journal of Molecular Medicine, 56, 154. https://doi.org/10.3892/ijmm.2025.5595
MLA
Li, A., Li, X., Wang, T., Song, J."SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption". International Journal of Molecular Medicine 56.4 (2025): 154.
Chicago
Li, A., Li, X., Wang, T., Song, J."SDF2 promotes glioma progression via GRP78‑mediated ERAD and copper homeostasis disruption". International Journal of Molecular Medicine 56, no. 4 (2025): 154. https://doi.org/10.3892/ijmm.2025.5595
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