PMCID string | Title string | Sentences string |
|---|---|---|
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Differential expression analysis was performed using the libraries DESeq2 version 1.36.0 (The R Foundation for Statistical Computing, Vienna, Austria) in R software version 4.2.1 (The R Foundation for Statistical Computing, Vienna, Austria). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | All volcano plots showed results of RNAseq as the statistical significance (adjusted p-value or q-value) versus the log2 FC and were generated using the libraries from ggplot2 version 3.3.36 in R software version 4.2.1. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Genes with an adjusted p-value (q-value) ≤ 0.01 and an absolute log 2-fold change (abs log2 FC) > 0.5 were considered to be statistically significant. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The ferroptosis database used for the analysis was retrieved on http://www.zhounan.org/ferrdb/ (accessed on 19 September 2022). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cell samples were lysed for 2 h at 4 °C in extraction buffer 5X PTR, purchased from Abcam (ab193970, Cambridge, United Kingdom), supplemented with extraction enhancer buffer 50X (Abcam, Cambridge, United Kingdom, Cat# ab193971) and EDTA-free protease inhibitor complete™ (Roche, Indianapolis, IN, USA, Cat# 11873580001). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The total protein concentration was measured based on the Bradford assay using the Bio-Rad Protein Assay Kit II (Bio-Rad, Hercules, CA, USA, Cat# 5000002). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Proteins were denatured in Laemmli sample buffer (Bio-Rad, Hercules, CA, USA, Cat# 1610747), heated at 95 °C for 5 min, separated by SDS-PAGE on 12% or 15% polyacrylamide gel, and transferred on a 0.2 μm nitrocellulose membrane (Bio-Rad, Hercules, CA, USA, Cat# 1620168). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Each membrane was blocked with Intercept blocking buffer PBS (Li-Cor, Lincoln, NE, USA, Cat# 927-70001). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Primary antibodies raised in different species and secondary antibodies, coupled with different fluorochromes (800 Li-Cor and 680 Li-Cor), were sequentially imaged on an Azure c500 imaging system (Azure Biosystems, Dublin, CA, USA) and displayed in green and red, respectively. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Image Lite Studio quantification software (version 5.2.5, LI-COR Biosciences, Lincoln, NE, USA) was used to quantify the fluorescence signals. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | A list of antibodies used is given in Table S6. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | All the whole western blot figures can be found in the Supplementary Materials. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Glutathione contents were measured using Glutathione Colorimetric Assay Kit (BioVision, CA, USA, Cat# K261). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | GIST882 cells were seeded at 500,000 cells/well and GIST48 cells at 300,000 cell/well in 6-well plates. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were treated with DMSO, CA3 (0.670 µM), or VP (2 µM for GIST882 cells and 0.5 µM for GIST48 cells) for 24 h and then harvested. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cell pellets were washed twice in ice-cold 0.01M PBS pH 7.4 and then lysed in ice-cold glutathione buffer for 20 min on ice. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cell lysates were centrifuged for 10 min at 13,000 rpm at 4 °C and each supernatant was transferred to a new tube. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | A small volume from the supernatant was taken for protein quantification. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Next, 5% sulfosalicylic acid (SSA) was added to the supernatant to precipitate and remove proteins from the sample and to protect GSH oxidation. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Each sample was then centrifuged at 8000× g for 10 min at 4 °C and the supernatant was transferred to a new tube for the next steps, according to the instructions in the kit. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The absorbance was read at 415 nm on a plate reader (iMark Microplate Absorbance Reader, BioRad, Hercules, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The content of GSH of each sample was calculated using a standard curve and normalized to the protein content in the lysate (µg GSH/mg protein). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Results are shown as scatter plots and are expressed as means ± SD of fold changes (treated samples normalized to untreated cells). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferrous iron was measured using the colorimetric QuantiChrom Iron Assay Kit (BioAssay Systems, CA, Cat# DIFE-250). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Briefly, cells were seeded in six-well plates (500,000 GIST882 cells or 300,000 GIST48 cells per well) 72 h before the start of the assay. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were treated for 24 h with VP or CA3 using concentrations mentioned above. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The next day, cells were washed with PBS and lysed on ice for 20 min in 55 µL of lysis buffer containing 1% Triton X-100 (PBS + 1% Triton X-100 + cOmplete EDTA-free protease inhibitor cocktail (Roche, Cat# 11873580001)). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cell lysates were centrifuged at 4 °C for 15 min at 13,000 rpm, and each supernatant was transferred to a new 1.5 mL tube. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | From the supernatant, 2 µL was taken aside for protein quantification and the remaining material was used for ferrous iron measurements following instructions of the kit. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The absorbance was read at 595 nm on a plate reader (iMark Microplate Absorbance Reader, BioRad, Hercules, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Ferrous iron concentration was calculated using a standard curve and normalized to the protein content of the lysate (µM/mg protein) for each sample. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Results are shown as scatter plots and are expressed as means ± SD of fold changes (treated samples normalized to untreated cells). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Flow cytometry analysis of C11-BODIPY green fluorescence was used as a readout for membrane lipid peroxidation, a hallmark of ferroptosis. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | When C11-BODIPY is oxidized, its emission fluorescence shifts from 591 nm (red) to 510 nm (green) . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cumene hydroperoxide (CumOOH), a strong oxidant, was used as a positive control for lipid peroxidation. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were seeded in a 6-well plate (TPP Techno Plastic Products AG, Trasadingen, Switzerland) at a cell density of 300,000 or 500,000 per well for GIST48 and GIST882 cells, respectively, supplemented with 2 mL of medium. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | After drug treatment, 1 µM of BODIPY™ 581/591 C11 (Invitrogen, OR, USA, Cat# D3861) was added to the well and incubated at 37 °C for 30 min. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were treated with either 50 µM of CumOOH for 45 min (GIST882 cells) or 100 µM for 30 min (GIST48) (Figures S1 and S2). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were washed twice with PBS 1x (Gibco, CA, USA, Cat# 10010-056) and trypsinized in trypsin-EDTA (0.25%) (Gibco, CA, USA, Cat# 25200-056). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were harvested and centrifuged for 2 min at 3000 rpm. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The pellet was resuspended in 500 µL of cold PBS/FBS 2%. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Hoechst 33342 (Invitrogen, OR, USA, Cat# H3570) was added prior to flow cytometry to exclude dead cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Flow cytometry was performed using a BD LSRFortessa™ X-20 cell analyzer (BD Bioscience). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Green fluorescence was detected using a 488 nm wavelength laser with a 505 nm dichroic mirror and a band-pass emission filter (530/30 nm), and blue fluorescence was detected using a 405 nm wavelength laser with a band-pass emission filter (450/50 nm). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Apoptosis was detected by flow cytometry using the FITC Active Caspase-3 Apoptosis Kit (BD Biosciences Pharmingen, SanDiego, CA and Lexington, KY, USA, Cat# 550480). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Briefly, GIST48 cells were seeded in 6-well plates at a cell density of 300,000 cells/ well. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were treated with 1 µM of staurosporine (STS) as a positive control for 4 h or with CA3 (0.670 µM) or VP (0.5 µM) for 24 h. Cells were collected, centrifuged, and resuspended in PBS. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Then, 250,000 cells per test were transferred to a new vial and then centrifuged. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Supernatants were removed and 100 µL of BD cytofix–cytoperm was added to each pellet for 30 min on ice. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were washed with BD Perm/Wash buffer (1x) and pelleted. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Supernatants were removed and caspase-3 active antibody was added to each tube at a concentration of 1/15 (diluted in BD Perm/Wash buffer 1x) for an incubation time of 1 h at room temperature. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were washed with BD Perm/Wash buffer (1x), pelleted, resuspended in PBS/FBS 2%, and analyzed by flow cytometry. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Flow cytometry was performed using a BD LSRFortessa™ X-20 cell analyzer (BD Bioscience). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Green fluorescence was detected using a 488 nm wavelength laser with a 505 nm dichroic mirror and a band-pass emission filter (530/30 nm). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Data from the flow cytometry experiment were saved in the proprietary *.fsc file format. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Analysis was performed using FlowJo 10.5.3 software (BD Biosciences, NJ, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Cells were selected for granularity (SSC-A x FSC-A) and singlets (FSC-W x FSC-H). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Living cells were selected (Hoechst x FSC-A) and represented as a FITC-A histogram. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Data were exported as CSV channel values. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | On the upper panel, Cumming estimation plots display all data points, presented as a swarm plot, and their distribution . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The number of events (single cells) analyzed is indicated in the figure. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The size effect (mean difference) is presented as a bootstrap sampling distribution and depicted as a dot with a 95% confidence interval (95% CI), indicated by the ends of the vertical error bars on the lower axis (Figures S1 and S2). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The mean difference in green fluorescence intensity of treated cells compared to untreated cells (DMSO) for multiple independent experimental replicates are shown as scatter plots ± SD, generated using Prism 9 software (GraphPad Software, Inc., La Jolla, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Estimation statistics were used for flow cytometry analysis to focus on the magnitude of the effect (effect size) and its precision. “ |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Data Analysis by Estimation Statistics” (DABEST) software (https://www.estimationstats.com/#/ (accessed on 22 July 2022)) was used as a Python script (https://github.com/ACCLAB/DABEST-python, accessed on 22 July 2022), running on a local PC to generate Cumming plots, for experiments that share one reference control group. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | For all other experiment, statistical analysis was performed with Prism 9 software (GraphPad Software, Inc., La Jolla, CA, USA), using ordinary one-way ANOVA, followed by Tukey’s test for ferrous iron and GSH measurements, the Pearson normality test, repeated-measures one-way ANOVA with Geisser–Greenhouse correction and Tukey’s multiple comparisons test for Western blot experiments, and multiple-ratio paired t-test for qPCR. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Contingency tables and Freeman–Halton extensions of Fisher’s test were performed to study the relationship between TFRC expression and non-continuous variables, such as tumor location, sex, mitotic figures, histological type, risk classification, YAP-ir, YAP activation, and KIT-ir. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The Kruskal–Wallis test was used to compare the mean of tumor size between the TFRC+++, TFRC++, and TFRC+ GIST samples. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | A p-value smaller than 0.05 was regarded as statistically significant for iron and GSH measurements and for Western blot quantifications. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | For the qPCR experiment, and for biological relevance, a higher stringency was applied and a p-value smaller 0.01 was regarded as statistically significant. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | All data are presented as mean ± SD. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Figures were prepared with Adobe Illustrator (Version 26.3.1, Adobe Inc., San Jose, CA, USA). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Imatinib-sensitive (GIST882) and imatinib-resistant (GIST48) cells were treated with class II FIN RSL3, a direct GPX4 inhibitor for 3 h. We first assessed the cell viability. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | RSL3 drastically reduced GIST882 and GIST48 cell viability (Figure 1A,B). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Liproxstatin (Lip), an antioxidant and ferroptosis inhibitor, protected GIST882 and GIST48 cells from RSL3 viability reduction. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We next investigated lipid peroxidation, the main hallmark of ferroptosis by flow cytometry using C11-BODIPY 581/591. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | RSL3 increased the mean difference in green fluorescence intensity (FITC-channel) after 3 h in a dose-dependent manner in both GIST882 and GIST48 cells (Figure 2A). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The antioxidant Lip (from 0.1 µM to 4 µM) completely abolished lipid peroxidation induced by RSL3 (150 nM) in both GIST882 and GIST48 cells (Figure 2B). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Deferoxamine (DFO), another ferroptosis inhibitor, acting through the chelation of iron, also prevented lipid peroxidation in both GIST882 and GIST48 cells (Figure 2C). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We conclude that RSL3 is a potent ferroptosis inducer in both GIST882 and GIST48 cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | To investigate a potential link between ferroptosis and YAP, cell viability was assessed in GIST882 and GIST48 cells after 24 h of treatment with the combination of VP, an inhibitor of YAP , and the two ferroptosis inhibitors, Lip and DFO. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We observed that Lip had a minor effect on viability reduction induced by VP in GIST882 and GIST48 cells, with a slight increase in IC50 values compared to the VP alone (Figure 3A). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Interestingly, DFO increased the viability of VP-treated GIST882 and GIST48 cells, as reflected by higher IC50 values (Figure 3B). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | These data indicate that ferroptosis plays a role in VP-induced cell death and that iron is important in this mechanism. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | VP is a photosensitizer used to treat macular degeneration and has been shown to inhibit YAP-TEAD interaction independently of light . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | However, ambient light can induce VP cytotoxicity through the formation of protein cross-linked oligomers and high-molecular-weight complexes or by ROS production, as suggested in the patient-derived xenograft of acute lymphoblastic leukemia . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Although all experiments with VP were sheltered from ambient light to minimize possible photoactivation, we also tested CA3, a compound with a chemical structure distinct from VP, recently coined as a YAP inhibitor . |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | To investigate the effect of CA3 on GIST882 and GIST48 cell viability after 24, 48, and 72 h, we performed viability assays (Figure 4A). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | CA3 drastically reduced viability in both cell lines at 24, 48, and 72 h, as shown by their respective IC50 values. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We then tested whether the antioxidant Lip and the iron chelator DFO reversed a reduction in viability induced by CA3 in GIST882 and GIST48 after 24 h (Figure 4B,C). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | Both Lip and DFO increased IC50 values when combined with CA3, as compared to CA3 alone. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | However, DFO was a more potent ferroptosis inhibitor than Lip to reverse the effect of CA3 on viability reduction. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | These data suggest that ferroptosis is the major cell death mechanism induced by CA3 in GIST882 and GIST48 cells. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We next assessed lipid peroxidation in both GIST882 and GIST48 cells (Figure 5, left and right panels). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We observed that VP increased the mean difference in green intensity fluorescence (FITC-channel) in GIST882 and GIST48 cells (Figure 5A) in a dose-dependent manner, corresponding to an increase in lipid peroxidation. |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The addition of different Lip and DFO concentrations halved the mean differences in green intensity fluorescence in GIST882 (Figure 5B,C, left panels) and GIST48 (Figure 5B,C, right panels). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | We next tested lipid peroxidation in GIST882 and GIST48 cells treated by CA3 for 24 h. CA3 also induced lipid peroxidation in a dose-dependent manner in GIST882 and GIST48 cells (Figure 5D). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The antioxidant Lip completely abolished lipid peroxidation in GIST882 and GIST48 cells (Figure 5E). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | The iron chelator DFO also prevented GIST882 and GIST48 from lipid peroxidation induced by CA3 (Figure 5F). |
PMC9599726 | Ferroptosis Induction and YAP Inhibition as New Therapeutic Targets in Gastrointestinal Stromal Tumors (GISTs) | These data suggest that VP largely, albeit not exclusively, induces cell death, whereas CA3 seems to exert its cytotoxicity entirely through ferroptosis. |
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