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PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The immunofluorescence result showed that STUB1 was present in the nucleus and cytoplasm, while GPX4 was predominantly located in the cytoplasm.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Furthermore, there was co-localization of STUB1 and GPX4 in the cytoplasm of both GIST-T1 (Pearson correlation coefficient = 0.728 ± 0.024) and GIST-882 cells (Pearson correlation coefficient = 0.689 ± 0.013; Figs. 4A and S4).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
First, we verified overexpression of STUB1 (Fig. 4B).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Next, we further explored whether STUB1 affected the protein level of GPX4, and found that STUB1 overexpression further reduced the protein level of GPX4 under IM treatment.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
However, STUB1 overexpression was no longer able to reduce the expression of GPX4 after treatment of the cells with the proteasome inhibitor MG132 (Fig. 4C).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Then we further explored whether STUB1 regulates GPX4 stability via the ubiquitination of GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
It was found that STUB1 overexpression led to an increase GPX4 degradation and prolonged the half-life of GPX4 protein in GIST-T1 cells (Fig. 4D).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Furthermore, STUB1 overexpression significantly increased GPX4 ubiquitination (Fig. 4E).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
To investigate whether STUB1 could directly regulate GPX4, protein-protein interaction was validated through a CO-IP assay using an anti-STUB1 and an anti-GPX4 antibody.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The results showed that STUB1 could co-precipitate with GPX4 (Figs. 4F and S5).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
In addition, proximity ligation assay (PLA) revealed a close proximity between STUB1 and GPX4 proteins and IM could significantly promote the proximity between the two proteins in GIST-T1 and 882 cells (P < 0.001; Fig. 4G).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
To further explore the mechanism by which STUB1 promoted the ubiquitination and degradation of GPX4, PhosphositePlus database (https://www.phosphosite.org/homeAction) was used to speculate that the K107, K126, K162, K167, and K191 sites were potential ubiquitination modified amino acid sites of GPX4, and RCSB Protein Data Bank (https://www.rcsb.org/) was used to simulate the corresponding point mutation positions (Figs. 4H and S6).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
We then constructed the point mutation plasmids by mutating the lysine (Lys) to the arginine (Arg), which cannot be modified by ubiquitination, and performed CO-IP experiments.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The results showed that among all the ubiquitination site mutants, only the K191R mutant had almost no detectable ubiquitination modification.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Compared with other mutants, the ubiquitination level of the GPX4-K191R mutant was downregulated significantly when it was co-transfected with STUB1 (Fig. 4I, J).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Computational molecular docking simulation analysis of STUB1 and GPX4 revealed a high possibility of combining structures (Fig. 4K).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
These findings collectively suggest that STUB1 is an E3 ligase of GPX4 that can promote its degradation by enhancing its ubiquitination.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Fig. 4STUB1 interacts with GPX4 to promote GPX4 ubiquitination in gastrointestinal stromal tumor (GIST).A Localization of STUB1 (green) and GPX4 (red) in GIST-T1 and 882 cells by confocal laser scanning immunofluorescence.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The nucleus is labeled by DAPI (blue).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Colocalization of STUB1 (green) and GPX4 (red) in cells appears yellow in the merged images.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Western blot analysis of STUB1 in GIST-T1 and 882 cells transfected with STUB1 overexpression plasmids or control plasmid.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C Western blotting was performed to assess the level of GPX4 in GIST-T1 cells transfected with GPX4 overexpression plasmids or control plasmid, and treated with IM (50 nM for T1 and 200 nM for 882) in the absence or presence of MG132 (5 μM) for 24 h. D STUB1 overexpression inhibited the stability of the GPX4 protein in GIST-T1 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The GPX4 protein levels were analyzed by western blotting after the cells were treated with CHX (100 μg/ml) for 0, 2, 4 or 8 h. E STUB1 overexpression promoted the ubiquitination of GPX4 protein in GIST-T1 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The ubiquitination of GPX4 protein was measured by CO-IP using an anti-ubiquitin antibody.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
F Direct interaction between STUB1 and GPX4 in GIST-T1 and 882 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Protein-protein interactions in GIST-T1 and 882 cells were validated by a CO-IP assay using an anti-STUB1 and an anti-GPX4 antibody.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
An antibody targeting IgG served as the negative control.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
G The representative images of proximity ligation assay (PLA).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
GIST-T1 and 882 cells were treated with IM (50 nM for T1 and 200 nM for 882) or a control vehicle for 24 h and then incubated with the anti-STUB1 and anti-GPX4 antibodies and detected by DuoLink probe (Olink Bioscience, Uppsala, Sweden).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
MG132 (5 μM) was added for stabilizing GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The red dots indicate the positive PLA signal.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
H The prediction of the lysine site of ubiquitination of GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
I The effects of the wild-type and different mutation sites of GPX4 on ubiquitin modification.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
J The effect of STUB1 on the ubiquitination modification of the K191 site of GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
K Computational molecular docking simulation analyses of STUB1 and GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Scale bar, 20 μm in A, 10 μm in G. Experiments were independently repeated three times.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Significance denoted by: ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
A Localization of STUB1 (green) and GPX4 (red) in GIST-T1 and 882 cells by confocal laser scanning immunofluorescence.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The nucleus is labeled by DAPI (blue).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Colocalization of STUB1 (green) and GPX4 (red) in cells appears yellow in the merged images.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Western blot analysis of STUB1 in GIST-T1 and 882 cells transfected with STUB1 overexpression plasmids or control plasmid.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C Western blotting was performed to assess the level of GPX4 in GIST-T1 cells transfected with GPX4 overexpression plasmids or control plasmid, and treated with IM (50 nM for T1 and 200 nM for 882) in the absence or presence of MG132 (5 μM) for 24 h. D STUB1 overexpression inhibited the stability of the GPX4 protein in GIST-T1 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The GPX4 protein levels were analyzed by western blotting after the cells were treated with CHX (100 μg/ml) for 0, 2, 4 or 8 h. E STUB1 overexpression promoted the ubiquitination of GPX4 protein in GIST-T1 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The ubiquitination of GPX4 protein was measured by CO-IP using an anti-ubiquitin antibody.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
F Direct interaction between STUB1 and GPX4 in GIST-T1 and 882 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Protein-protein interactions in GIST-T1 and 882 cells were validated by a CO-IP assay using an anti-STUB1 and an anti-GPX4 antibody.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
An antibody targeting IgG served as the negative control.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
G The representative images of proximity ligation assay (PLA).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
GIST-T1 and 882 cells were treated with IM (50 nM for T1 and 200 nM for 882) or a control vehicle for 24 h and then incubated with the anti-STUB1 and anti-GPX4 antibodies and detected by DuoLink probe (Olink Bioscience, Uppsala, Sweden).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
MG132 (5 μM) was added for stabilizing GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The red dots indicate the positive PLA signal.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
H The prediction of the lysine site of ubiquitination of GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
I The effects of the wild-type and different mutation sites of GPX4 on ubiquitin modification.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
J The effect of STUB1 on the ubiquitination modification of the K191 site of GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
K Computational molecular docking simulation analyses of STUB1 and GPX4.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Scale bar, 20 μm in A, 10 μm in G. Experiments were independently repeated three times.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Significance denoted by: ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
We evaluated the potential tumor suppressive effect of RSL3 in combination with IM in GIST-T1 and GIST-882 cells by treating with different concentrations of drug combinations.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Our data revealed that a single treatment with IM or RSL3 significantly inhibited the GIST cell activity (Fig. 5A).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The synergistic antitumor effect of RSL3 and IM was analyzed using CompuSyn software, which applies the median effect theorem of the law of mass action and its combination index theorem to facilitate pharmacodynamic studies and computerized analysis simulations.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Combination index (CI) values were calculated based on the drug combination principles, as proposed by Chou—Talalay (CI <1, =1, and >1 indicate synergic, additive or antagonistic effects, respectively) .
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Our results demonstrated that the CI values in both GIST-T1 and GIST-882 cells were <1, implying that RSL3 exerted synergistic effects with IM in both GIST-T1 and GIST-882 cells, offering an advantage of the combination treatment (Fig. 5B).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The GSH/GSSG ratio (Fig. 5C) and the expression of GPX4 (Fig. 5D, E) in RSL3 combined with the IM group were lower than those in IM or RSL3 group alone.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Thus, the combination of RSL3 and IM could synergistically inhibit GIST cell viability.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Fig. 5Combination treatment of Imatinib (IM) and RSL3 for gastrointestinal stromal tumor (GIST) in vitro.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
A CCK-8 viability assay following treatment for 24 h with escalating concentrations of IM (0, 25, 50, 100, and 200 nM for T1 and 0, 50, 100, 200, and 400 nM for 882) and RSL3 (0, 0.5, 1, and 2 μM for T1 and 0, 0.25, 0.5 and 1 μM for 882) in GIST-T1 and 882 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Synergy was calculated using CompuSyn and a combination index value of under 1.0 was considered synergy.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C The relative GSH/GSSG ratios were assayed after GIST-T1 and 882 cells were treated with IM (50 nM for T1 and 200 nM for 882) and RSL3 (2 μM for T1 and 1 μM for 882) for 24 h. D, E Western blotting analysis of SLC7A11 and GPX4 protein expression after combination treatment of IM and RSL3 in GIST-T1 (D) and 882 cells (E) for 24 h. Experiments were independently repeated three times.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Significance denoted by: ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
A CCK-8 viability assay following treatment for 24 h with escalating concentrations of IM (0, 25, 50, 100, and 200 nM for T1 and 0, 50, 100, 200, and 400 nM for 882) and RSL3 (0, 0.5, 1, and 2 μM for T1 and 0, 0.25, 0.5 and 1 μM for 882) in GIST-T1 and 882 cells.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Synergy was calculated using CompuSyn and a combination index value of under 1.0 was considered synergy.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C The relative GSH/GSSG ratios were assayed after GIST-T1 and 882 cells were treated with IM (50 nM for T1 and 200 nM for 882) and RSL3 (2 μM for T1 and 1 μM for 882) for 24 h. D, E Western blotting analysis of SLC7A11 and GPX4 protein expression after combination treatment of IM and RSL3 in GIST-T1 (D) and 882 cells (E) for 24 h. Experiments were independently repeated three times.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Significance denoted by: ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The GIST-T1 cells were subcutaneously implanted into the backs of immunodeficient nude mice.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
When the tumor volume reached 50 mm, the mice were randomly divided into four groups and subsequently treated with either DMSO, IM, RSL3 or a combination of IM and RSL3 (Fig. 6A).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
No significant changes in body weight or signs of toxicity were observed in any of the treatment groups (Fig. 6B).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Compared to the DMSO group, tumor growth was inhibited in both the IM and RSL3 groups, with the most significant inhibition observed in the IM combined with RSL3 group (Fig. 6C and S7).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
In addition, Western blotting analysis of KIT, STUB1, GPX4, Ki-67, and 4-HNE protein levels was performed on subcutaneous xenograft tumors.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The results showed that the expression of KIT, Ki-67, and GPX4 were decreased and the expression of STUB1 and 4-HNE was increased in the IM-treated GIST-T1 xenografts, indicating that IM could induce ferroptosis in vivo.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
After treatment with RSL3, the expression of GPX4 and Ki-67 was decreased and the expression of 4-HNE was increased.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
In addition, the expression of Ki-67 and GPX4 was the lowest and the expression of 4-HNE was the highest in the combined-treated GIST-T1 xenograft model (Figs. 6D–F and S8).
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
The representative IHC staining pictures of hematoxylin-eosin (HE), KIT, STUB1, GPX4, Ki-67, and 4-HNE are presented in Fig. 6G.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
These findings suggest that RSL3 enhances IM-induced lipid ROS and ferroptosis in GIST, leading to synergistic inhibition of GIST in vivo.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Fig. 6Subcutaneous tumor models of gastrointestinal stromal tumor (GIST) were established to analyze the synergy effect of Imatinib (IM) and RSL3.A GIST-T1 cells were subcutaneously transplanted into the dorsolateral side of nude mice.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
When the subcutaneous tumor volume reached 50 mm, the mice were randomly divided into four groups (six mice in each group), including the vehicle (DMSO), IM (100 mg/kg), RSL3 (10 mg/kg) or IM combined with RSL3 treatment groups, and then were injected intraperitoneally every there day.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
All mice were euthanized 18 days later.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Body weight was measured every 3 days until day 18.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C The images and quantitative analysis of tumor volume on day 18 after treatment.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
D–F Western blot analysis of the protein levels of Ki-67, KIT, STUB1, GPX4, and 4-HNE in GIST cell xenograft tumor tissues of mice from vehicle (DMSO), IM, RSL3, and combination treatment groups.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
n = 3 mice per group.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
G Immunohistochemical staining representative pictures of hematoxylin-eosin (HE), KIT, STUB1, GPX4, Ki-67, and 4-HNE in GIST cell xenograft tumor tissues of mice from vehicle (DMSO), IM, RSL3, and combination treatment groups.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Scale bar, 100 μm.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
DMSO dimethyl sulfoxide, IM Imatinib, RSL3 Ras-selective lethal small molecule 3, 4-HNE 4-hydroxy-2-nonenal.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
Significance denoted by: ns not significant, *P < 0.05, **P < 0.01, and ***P < 0.001.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
A GIST-T1 cells were subcutaneously transplanted into the dorsolateral side of nude mice.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
When the subcutaneous tumor volume reached 50 mm, the mice were randomly divided into four groups (six mice in each group), including the vehicle (DMSO), IM (100 mg/kg), RSL3 (10 mg/kg) or IM combined with RSL3 treatment groups, and then were injected intraperitoneally every there day.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
All mice were euthanized 18 days later.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
B Body weight was measured every 3 days until day 18.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
C The images and quantitative analysis of tumor volume on day 18 after treatment.
PMC10728200
Imatinib induces ferroptosis in gastrointestinal stromal tumors by promoting STUB1-mediated GPX4 ubiquitination
D–F Western blot analysis of the protein levels of Ki-67, KIT, STUB1, GPX4, and 4-HNE in GIST cell xenograft tumor tissues of mice from vehicle (DMSO), IM, RSL3, and combination treatment groups.