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Article Open Access

HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis

  • Authors:
    • Wenwen Han
    • Hongli Chen
    • Bomiao Cui
    • Jiao Chen
    • Ping Zhang
    • Yun Feng
  • View Affiliations / Copyright

    Affiliations: State Key Laboratory of Oral Diseases and National Clinical Research Centre for Oral Diseases and West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, P.R. China
    Copyright: © Han 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: January 22, 2026
       https://doi.org/10.3892/mmr.2026.13807
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Abstract

High‑mobility group nucleosomal‑binding domain 2 (HMGN2) is an abundant conserved protein that acts as a non‑histone nuclear DNA‑binding protein. HMGN2 can be released by activated peripheral blood mononuclear cells, CD8+ T cells and γδ T cells, and can induce tumour cell apoptosis. In the present study, receptors of HMGN2 were detected on tumour cell membranes and the mechanism by which HMGN2 induces tumour cell apoptosis was examined. Flow cytometry was used to determine the degree of HMGN2‑induced apoptosis. To identify notable HMGN2 receptors on tumour cells, the present study used immunoprecipitation and mass spectrometry (IP/MS) to identify protein complexes. Western blotting and immunofluorescence were used to confirm interactions between HMGN2 and oligosaccharyltransferase subunit STT3B (STT3B), and to elucidate the downstream regulatory mechanism of HMGN2. The predictive tools ZDOCK and AlphaFold3 were used to determine the binding conformation of HMGN2 to STT3B. HMGN2 was shown to bind to the membrane and induce the apoptosis of CAL‑27 tumour cells. STT3B was identified via IP/MS as a receptor of HMGN2 on the CAL‑27 membrane and subsequently identified as an important receptor of HMGN2 via an anti‑STT3B blocking assay. ZDOCK and AlphaFold3 analyses revealed that HMGN2 and STT3B formed a stable protein docking model. After incubation with HMGN2, the expression of programmed cell death 1 ligand 1 (PD‑L1)/caspase‑1/gasdermin D (GSDMD) axis components was significantly increased, and PD‑L1 was translocated into the nucleus from the membrane of CAL‑27 cells. The results of the present study indicated that extracellular HMGN2 induced pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis.

View Figures

Figure 1

HMGN2 induces apoptosis in CAL-27
cells. (A) CAL-27 cells were observed under a bright-field
microscope after 24 h of incubation with (Aa) 0, (Ab) 5, (Ac) 10 or
(A-d) 20 µg/ml HMGN2. (B) Representative flow cytometry plots
showing apoptosis of CAL-27 cells following incubation with 20
µg/ml HMGN2. (C) Relative percentage of apoptotic CAL-27 cells
treated with HMGN2 for (Ca) 12 and (Cb) 24 h. The control group was
treated with PBS. Annexin+ PI− represents
cells in the early phase of apoptosis and Annexin+
PI+ represents cells in the last phase of apoptosis.
Scale bar, 300 µm. Data are presented as the mean ± SEM (n=3).
##P<0.01 vs. control. Ctrl, control; HMGN2,
high-mobility group nucleosomal-binding domain 2; ns, not
significant; PI, propidium iodide.

Figure 2

Microscopic imaging of CAL-27 cells
after incubation with FITC-labelled HMGN2. HMGN2 was translocated
into CAL-27 cells through binding to the cell membrane after
incubation with 20 µg/ml FITC-labelled HMGN2 for 10 min, 1 or 2 h.
Scale bar, 20 µm. Ctrl, control; HMGN2, high-mobility group
nucleosomal-binding domain 2.

Figure 3

Predicted receptors of HMGN2
determined by IP/MS. (A) IP/MS analysis of HMGN2-associated
proteins. The bead-peptide eluate was resolved by SDS-PAGE and
Coomassie blue staining. (B) Numbers of proteins identified via
IP/MS from magnetic bead-bound antibody-protein complexes and via
SDS-PAGE gel separation. (C) Subcellular localization of STT3B in
CAL-27 cells. Scale bar, 10 µm. IP, immunoprecipitation; MS, mass
spectrometry; TP, total protein; MP, membrane protein; HMGN2,
high-mobility group nucleosomal-binding domain 2; STT3B,
oligosaccharyltransferase subunit STT3B.

Figure 4

STT3B is an important receptor of
HMGN2 on CAL-27 cell membranes. (A and B) CAL-27 cells were
incubated with 20 µg/ml HMGN2 (experimental group) or with PBS
(control group). Co-IP was carried out using anti-HMGN2 or
anti-STT3B antibodies against (A) MP and (B) TP. Immunocomplexes
were subsequently detected by IB using anti-STT3B or anti-HMGN2
antibodies. (C) CAL-27 cells were observed under a fluorescence
microscope after depletion of STT3B using 10 µg/ml anti-STT3B
antibody. Scale bar, 20 µm. (D) Rigid protein-protein docking
prediction for the interaction between STT3B and HMGN2. (E)
Interaction structure of STT3B and HMGN2 simulated in AlphaFold3.
Ctrl, control; IB, immunoblotting; IP, immunoprecipitation; TP,
total protein; MP, membrane protein; STT3B,
oligosaccharyltransferase subunit STT3B; HMGN2, high-mobility group
nucleosomal-binding domain 2.

Figure 5

HMGN2 induces pyroptosis in CAL-27
cells through the PD-L1/caspase-1/GSDMD axis. (A) Representative
western blot analysis of the PD-L1/caspase-1/GSDMD axis, and the
HMGN2 and STT3B proteins in CAL-27 cells after incubation with 20
µg/ml HMGN2 for 10 or 60 min. The Ctrl group was treated with PBS.
(B) Semi-quantitative analyses of the PD-L1/caspase-1/GSDMD axis,
and the HMGN2 and STT3B proteins in CAL-27 cells (n=3). (C)
Representative images showing PD-L1 translocation in CAL-27 cells
treated with HMGN2. The Ctrl group was treated with PBS. Scale bar,
20 µm. Data are presented as the mean ± SEM. *P<0.05 and
**P<0.01; #P<0.05 and ##P<0.01.
Ctrl, control; ns, not significant; HMGN2, high-mobility group
nucleosomal-binding domain 2; PD-L1, programmed cell death 1 ligand
1; GSDMD-N, N-terminal domain of gasdermin D; GSDMD-FL, full-length
gasdermin D; STT3B, oligosaccharyltransferase subunit STT3B;
C-caspase-1, cleaved caspase-1.
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Copy and paste a formatted citation
Spandidos Publications style
Han W, Chen H, Cui B, Chen J, Zhang P and Feng Y: <p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>. Mol Med Rep 33: 97, 2026.
APA
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., & Feng, Y. (2026). <p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>. Molecular Medicine Reports, 33, 97. https://doi.org/10.3892/mmr.2026.13807
MLA
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., Feng, Y."<p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>". Molecular Medicine Reports 33.3 (2026): 97.
Chicago
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., Feng, Y."<p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>". Molecular Medicine Reports 33, no. 3 (2026): 97. https://doi.org/10.3892/mmr.2026.13807
Copy and paste a formatted citation
x
Spandidos Publications style
Han W, Chen H, Cui B, Chen J, Zhang P and Feng Y: <p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>. Mol Med Rep 33: 97, 2026.
APA
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., & Feng, Y. (2026). <p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>. Molecular Medicine Reports, 33, 97. https://doi.org/10.3892/mmr.2026.13807
MLA
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., Feng, Y."<p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>". Molecular Medicine Reports 33.3 (2026): 97.
Chicago
Han, W., Chen, H., Cui, B., Chen, J., Zhang, P., Feng, Y."<p>HMGN2 induces pyroptosis in tumour cells by modulating the STT3B/PD‑L1/caspase‑1/GSDMD axis</p>". Molecular Medicine Reports 33, no. 3 (2026): 97. https://doi.org/10.3892/mmr.2026.13807
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