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ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells

  • Authors:
    • Xuhao Chen
    • Mingqi Luo
    • Xiaoyu Niu
    • Wang Wang
    • Huanhuan Cao
    • Liwei Zhang
    • Ruolun Wei
    • Ping Duan
  • View Affiliations / Copyright

    Affiliations: Department of Pathophysiology, College of Medicine, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Department of Physiology and Neurobiology, College of Medicine, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Department of Neurosurgery, Stanford University, Stanford, CA 94305, USA
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 97
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    Published online on: September 24, 2025
       https://doi.org/10.3892/ijo.2025.5803
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Abstract

Ferroptosis is an iron‑dependent type of regulated cell death which is dysregulated in several tumors, including glioblastoma (GBM). Zinc finger and BTB domain‑containing protein 20 (ZBTB20), a transcription repressor, is expressed at low levels in GBM and suppresses GBM cell proliferation through the ERK signaling pathway. However, the effect of ZBTB20 on ferroptosis has not been explored. The present study aimed to explore the role of ZBTB20 in ferroptosis of glioma cells and its underlying mechanism. The present study demonstrated that both ZBTB20 expression and ferroptosis levels in GBM cells were lower than that in normal glial cells. Gain‑ and loss‑of‑function experiments revealed that ZBTB20 overexpression promoted ferroptosis and ZBTB20 knockdown inhibited ferroptosis in GBM cells. Moreover, the results demonstrated that ZBTB20 transcriptionally repressed the expression of transmembrane protein 109 (TMEM109) in GBM cells, assessed using dual‑luciferase reporter and chromatin immunoprecipitation assays. TMEM109 is mainly localized on the endoplasmic reticulum (ER) membrane of cells and regulates calcium leakage at the ER or sarcoplasmic reticulum. The present study revealed that TMEM109 overexpression inhibited ferroptosis and TMEM109 knockdown promoted ferroptosis in GBM cells. Using co‑transfection experiments, it was further revealed that the promotive effect of ZBTB20 can reverse the inhibitory effect of TMEM109 on ferroptosis. In conclusion, the findings indicated that ZBTB20 promotes ferroptosis in GBM cells through transcriptionally repressing the expression of TMEM109.
View Figures

Figure 1

ZBTB20 expression and ferroptosis
level in GBM cells. (A-E) Expression of ZBTB20, TMEM109, SLC7A11
and GPX4 in U251 and HA1800 cells detected by (A) western blotting
and (B-E) the corresponding histograms. (F-J) Expression of ZBTB20,
TMEM109, SLC7A11 and GPX4 in U87MG and HA1800 cells detected by (F)
western blotting and (G-J) the corresponding histograms. (K and L)
ROS level detected by fluorescence microscopy and the corresponding
histogram. (M) Histogram of Fe2+ content. (N) Overall
survival of ZBTB20. (O) Overall survival of TMEM109. Data are shown
as the mean ± standard deviation, n=3. *P<0.05,
**P<0.01 and ***P<0.001. ZBTB20, zinc
finger and BTB domain containing 20, GBM, glioblastoma; TMEM109,
transmembrane protein 109; SLC7A11, solute carrier family 7 member
11, GPX4, glutathione peroxidase 4; ROS, reactive oxygen
species.

Figure 2

ZBTB20 promotes ferroptosis in U251
GBM cells. (A-D) Expression of ZBTB20, SLC7A11 and GPX4 detected by
(A) western blotting and (B-D) the corresponding histograms in GBM
cells with ZBTB20 overexpression. (E-H) Expression of ZBTB20,
SLC7A11 and GPX4 detected by (E) western blotting and (F-H) the
corresponding histograms in GBM cells with ZBTB20 knockdown. (I-K)
ROS levels detected by (I) fluorescence microscopy and the
corresponding histogram in GBM cells with ZBTB20 (J) overexpression
and (K) knockdown. (L and M) Histograms of Fe2+ content
in GBM cells with ZBTB20 (L) overexpression and (M) knockdown.
*P<0.05, **P<0.01 and
***P<0.001. ZBTB20, zinc finger and BTB domain
containing 20, GBM, glioblastoma; TMEM109, transmembrane protein
109; SLC7A11, solute carrier family 7 member 11, GPX4, glutathione
peroxidase 4; ROS, reactive oxygen species.

Figure 3

ZBTB20 transcriptionally represses
TMEM109 expression in U251 GBM cells. (A-D) Expression of (A and B)
ZBTB20 and (C and D) TMEM109 in GBM cells with ZBTB20
overexpression detected by reverse transcription-quantitative PCR
with the corresponding histogram. (E-H) Expression of ZBTB20 and
TMEM109 in GBM cells with ZBTB20 knockdown detected by (E and G)
western blotting with (F and H) the corresponding histogram. (I)
Results of chromatin immunoprecipitation assay. (J) Results of the
dual-luciferase reporter gene experiments. *P<0.05,
**P<0.01 and ***P<0.001. ZBTB20, zinc
finger and BTB domain containing 20, GBM, glioblastoma; TMEM109,
transmembrane protein 109.

Figure 4

TMEM109 represses ferroptosis in U251
GBM cells. (A-D) Expression of ZBTB20, SLC7A11 and GPX4 detected by
(A) western blotting and (B-D) the corresponding histograms in GBM
cells with TMEM109 overexpression. (E-H) Expression of ZBTB20,
SLC7A11 and GPX4 detected by (E) western blotting and (F-H) the
corresponding histograms in GBM cells with TMEM109 knockdown. (I-K)
ROS levels detected by (I) fluorescence microscopy and the
corresponding histogram in GBM cells with TMEM109 (J)
overexpression and (K) knockdown. (L and M) Histograms of
Fe2+ content in GBM cells with TMEM109 (L)
overexpression and (M) knockdown. *P<0.05,
**P<0.01 and ***P<0.001. ZBTB20, zinc
finger and BTB domain containing 20, GBM, glioblastoma; TMEM109,
transmembrane protein 109; SLC7A11, solute carrier family 7 member
11, GPX4, glutathione peroxidase 4; ROS, reactive oxygen
species.

Figure 5

ZBTB20 promotes ferroptosis through
repressing TMEM109 expression in U251 GBM cells. (A-D) Expression
of ZBTB20, TMEM109, SLC7A11 and GPX4 detected by reverse
transcription-quantitative PCR and the corresponding histograms in
GBM cells with ZBTB20 and/or TMEM109 overexpression. (E-I)
Expression of ZBTB20, TMEM109, SLC7A11 and GPX4 detected by (E)
western blotting and (F-I) the corresponding histograms in GBM
cells with ZBTB20 and/or TMEM109 overexpression. (J and K) ROS
levels detected by (J) fluorescence microscopy and (K) the
corresponding histogram. (L) Histograms of Fe2+ content
in GBM cells with ZBTB20 and/or TMEM109 overexpression. (M)
Histograms of MDA content in GBM cells with ZBTB20 and/or TMEM109
overexpression. *P<0.05, **P<0.01 and
***P<0.001. ZBTB20, zinc finger and BTB domain
containing 20, GBM, glioblastoma; TMEM109, transmembrane protein
109; SLC7A11, solute carrier family 7 member 11, GPX4, glutathione
peroxidase 4; ROS, reactive oxygen species; MDA,
malondialdehyde.
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Copy and paste a formatted citation
Spandidos Publications style
Chen X, Luo M, Niu X, Wang W, Cao H, Zhang L, Wei R and Duan P: ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells. Int J Oncol 67: 97, 2025.
APA
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L. ... Duan, P. (2025). ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells. International Journal of Oncology, 67, 97. https://doi.org/10.3892/ijo.2025.5803
MLA
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L., Wei, R., Duan, P."ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells". International Journal of Oncology 67.6 (2025): 97.
Chicago
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L., Wei, R., Duan, P."ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells". International Journal of Oncology 67, no. 6 (2025): 97. https://doi.org/10.3892/ijo.2025.5803
Copy and paste a formatted citation
x
Spandidos Publications style
Chen X, Luo M, Niu X, Wang W, Cao H, Zhang L, Wei R and Duan P: ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells. Int J Oncol 67: 97, 2025.
APA
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L. ... Duan, P. (2025). ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells. International Journal of Oncology, 67, 97. https://doi.org/10.3892/ijo.2025.5803
MLA
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L., Wei, R., Duan, P."ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells". International Journal of Oncology 67.6 (2025): 97.
Chicago
Chen, X., Luo, M., Niu, X., Wang, W., Cao, H., Zhang, L., Wei, R., Duan, P."ZBTB20 promotes ferroptosis through inhibiting TMEM109 expression in glioblastoma cells". International Journal of Oncology 67, no. 6 (2025): 97. https://doi.org/10.3892/ijo.2025.5803
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