PMCID string | Title string | Sentences string |
|---|---|---|
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The results showed that inducing ferroptosis enhanced the sensitivity of resistant cells to imatinib. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Furthermore, we found that combinative treatment with imatinib and β‐elemene can hyperactivate HMOX1 and eventually lead to ferroptosis. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Moreover, by thermal proteome profiling (TPP), we identified N6AMT1 as the target protein of β‐elemene. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Mechanistically, β‐elemene deactivates N6AMT1 and then induces ferroptosis via the nuclear factor erythroid 2‐related factor 2 (NRF2)‐HMOX1 signalling axis. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Taken together, our work therefore provides a new therapeutic strategy for imatinib resistance in patients with advanced GIST. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The GIST cell line GIST‐882 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST‐T1 was a gift from Professor Haibo Qiu (Sun Yat‐sen University Cancer Center [SYSUCC]), cultured as described in previous literature. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST‐T1and GIST‐882 cells were cultured in RPMI1640 medium supplemented with 15% fetal bovine serum, 1% penicillin/streptomycin, 1% l‐glutamine and 2 µg/mL Gentamycin. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | To establish cells resistant to imatinib, GIST‐882 and GIST‐T1 cells were exposed to gradually increasing concentrations of imatinib as previously described. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST cell lines were generated through at least 6 months of drug resistance. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The administration of the specific imatinib concentration was continued until the cells grew normally. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Subsequently, the concentration was increased, and the above process was repeated to obtain imatinib‐resistant cell lines through repeated induction. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The cell counting kit‐8 (CCK‐8) assay was used to evaluate the half‐maximal inhibitory concentration (IC50) of GIST cell lines. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | For the functional assays, imatinib was removed for at least 1 week to avoid acute effects. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | All cell lines were cultured in an incubator with 37 ° C and 5% CO2. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Ferroptosis activators erastin (HY‐15763) and RAS‐selective lethal 3 (RSL3; HY‐100218) were purchased from MedChemExpress. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Ferroptosis inhibitor ferrostatin‐1 (Fer‐1, HY‐100579) was purchased from MedChemExpress. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Hemin (51280) and zinc protoporphyrin‐9 (ZnPP, MB4231) were purchased from Sigma‐Aldrich and Meilunbio, respectively. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cell viability was evaluated using the CCK‐8 (LJ621, Dojindo) according to the product manual. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | A total of 2000 GIST cells were seeded in 96‐well plates and treated with corresponding processes. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After the cells were treated with drugs, the medium was replaced with 100 µL fresh medium containing 10 µL CCK‐8 reagent. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After incubation for 2 h, the absorbance was detected at a wavelength of 450 nm, and growth curves were generated to determine the cell inhibition rate. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | To calculate IC50 values for all drugs, GraphPad Prism Software was used. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | We utilised the CompuSyn software (ComboSyn, Inc.) to assess the drug synergism effect, which involves the computation of the combination index (CI). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | CI values offer quantitative definitions for additive effect (CI = 1), synergism (CI < 1), and antagonism (CI > 1). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cell death was measured by propidium iodide (PI; SIGMA, Cat#537059) staining. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were seeded in six‐well plates 1 day before treatment with drugs. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | On the next day, cells were treated with the indicated drugs for 48 h, all cells including the floating cells in the culture medium were harvested by trypsinisation and resuspended in fresh PBS containing PI. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After incubation for 10 min, the cells were analysed by flow cytometry. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Data were analysed using FlowJo software. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were seeded in six‐well plates (2000 cells per well) with three replicates. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Following cell adhesion, the medium was replaced with a medium containing the indicated drugs. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After 14 days with a complete medium at 37°C with 5% CO2. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were then fixed with methanol and stained with crystal violet for 30 min at 25°C, washed with ddH2O and the colonies of each well were counted. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells and tissues were lysed in a lysis buffer (50 mM TrisHCl, pH 7.4, 150 mM NaCl, 1% NP‐40 [Beyotime, P0013F], .1% SDS, .5% sodium deoxycholate [Sigma‐Aldrich, 302‐954], 1 mM EDTA and 10% glycerol) and centrifuged at 13 000 × g for 30 min at 4°C. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Protein concentration was determined using a BCA Protein Assay Kit (Thermo Fisher Scientific). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Equal amounts of total protein lysates were separated on 10% SDS‐PAGE and transferred to a PVDF membrane (Bio‐Rad). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Subsequently, the membranes were blocked with 5% non‐fat milk at room temperature for 1 h, followed by an overnight incubation at 4°C with primary antibodies. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Primary antibodies used in this study are anti‐caspase‐3 (Proteintech, 19677‐1‐AP), anti‐cleaved caspase‐3 (CST, 9664S), anti‐caspase‐7 (Proteintech, 27155‐1‐AP), anti‐cleaved caspase‐7 (CST, 9491T), anti‐GSDMD (Proteintech, 20770‐1‐AP), anti‐GSDME (Proteintech, 13075‐1‐AP), anti‐glutathione peroxidase 4 (GPX4; Affinity Biosciences, DF6701), anti‐ferritin heavy chain (FTH1; Affinity Biosciences, DF4828), anti‐ACSL3 (Affinity Biosciences, DF9606), anti‐vinculin (CST, 13901S), anti‐HMOX1 (Proteintech, 10701‐1‐AP), anti‐NRF2 (Proteintech, 16396‐1‐AP), anti‐N6AMT1 (Proteintech, 16211‐1‐AP), anti‐SLC7A11 (Proteintech, 26864‐1‐AP), anti‐FSP1 (Proteintech, 20886‐1‐AP), anti‐dihydroorotate dehydrogenase (DHODH; Proteintech, 14877‐1‐AP) and anti‐SMYD2 (Proteintech, 21290‐1‐AP). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The PVDF membrane was washed with TBST buffer and incubated with the secondary antibodies labelled with HRP‐conjugated secondary antibodies (Neobioscience) for 2 h at room temperature. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Protein signals were detected with an enhanced chemiluminescent kit (Thermo Scientific). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Paraffin‐embedded GIST samples were collected from 40 patients who underwent surgical therapy at SYSUCC. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Written consent was obtained from the participating patients before surgery, and the clinical and histopathological data provided to researchers were anonymised. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The study was approved by the Institutional Research Ethics Committee of SYSUCC (Guangzhou, China, Approval Number: B2022‐465‐01). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | For IHC staining, paraffin‐embedded GIST specimens were performed as described previously. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Primary antibodies used in this study are anti‐GPX4 (Affinity Biosciences, DF6701), anti‐FTH1 (Affinity Biosciences, DF4828), anti‐Ki 67 (Abcam, ab15580), anti‐HMOX1 (Proteintech, 10701‐1‐AP), anti‐NRF2 (Proteintech, 16396‐1‐AP), anti‐4‐hydroxynonenal (R&D, MAB3249). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | IHC scores were recorded as the degree of staining intensity and percentage of positive cells. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The staining extent that scored according to the percentage of positively stained cells ranged from 0 to 3 (0, 0%–5%; 1, 5%–25%; 2, 26%–50%; 3, 51%–75%; and 4, 76%–100%), although the intensity of staining was scored as 0 (negative staining), 1 (weak staining), 2 (moderate staining), and 3 (strong staining). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Final IHC staining scores were evaluated by two independent gastrointestinal pathologists blinded to the patient's clinical characteristics. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were inoculated in culture dishes with a cell density of 1 × 10/mL and treated according to designated groups for 24 h. The cells were incubated with 1 µmol/L FerroOrange working solution (Dojindo) for 30 min and then observed under a fluorescence microscope. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | All images were acquired with the same instrument parameters and processed with the same settings to maximise the ability to compare results between conditions. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were seeded at a density of 1 × 10cells per 60 mm culture dish and treated according to designated groups. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The relative concentration of MDA was measured using the MDA assay kit (Dojindo) according to the manufacturer's instructions. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Protein concentration was assayed using a Thermo Fisher Scientific BCA Protein Assay Kit according to the manufacturer's instructions. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Dichlorodihydrofluorescein diacetate (DCFH‐DA) assay kit (Beyotime) was used to detect intracellular ROS production. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Cells were seeded in six‐well plates and treated with the indicated drugs. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Then, cells were incubated with DCFH–DA at a final concentration of 10 µM in medium without FBS at 37°C for 30 min and washed three times with medium. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The level of ROS was determined by flow cytometer. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The level of ROS generation was analysed with FlowJo software (FlowJo). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Lipid peroxidation was detected using C11 BODIPY581/591 (Thermo Fisher Scientific). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST cells (1 × 10 cells per dish) were plated in culture dishes and incubated for the indicated drugs for 24 h. After treatment, the cells were incubated in a medium containing 5 µM BODIPY 581/591 C11 at 37°C for 30 min for live‐cell imaging. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The cells were imaged at 40 × magnification using a Leica microscope to detect the oxidised forms of the probe. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | All images were acquired with the same instrument parameters and processed with the same settings to maximise the ability to compare results between conditions. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST cells were seeded in dishes and subjected to different treatments. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After 24 h, cells were incubated with a pre‐chilled 2% glutaraldehyde solution for 2 h at 4°C to fix the cell pellet. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The cells were stained with 2% uranyl acetate solution for 2 h and then dehydrated in 50%, 70%, 90%, 95% and 100% acetone. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The cells were embedded in Spurr embedding kit (KYD bio, 14300), and ultrathin sections were prepared for observation under an electron microscope (HITACHI). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The small interfering RNA (siRNA) targeting N6AMT1 and SMYD2 were synthesised by GENE CREATE. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Transfection was conducted using jetPRIME reagent (Polyplus, 101000046) as recommended. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The siRNA sequences used for gene knockdown in this study were listed in Supporting Information Table S1. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | After transfection for 8 h, the medium was refreshed with complete medium, and these cells were cultivated overnight in an incubator before being re‐seeded for experiments. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | For overexpression of HMOX1 and N6AMT1, negative control or over‐expressing plasmids were co‐transfected into HEK293T with pHelper and pEnv. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The virus was harvested from the transfected HEK293T cells, and the GIST cells were then infected with the viral supernatants for 2 consecutive days. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Stable GIST cell lines were selected by treating with puromycin (2 µg/mL) for 10 days. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The total RNA was extracted using TRIzol reagent (Invitrogen, 15596026) and adjusted to 200 µg/mL. .5 µg total RNA was used for reverse transcription. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Complementary DNA was generated using the PrimeScript RT reagent kit (TaKaRa Bio, RR037A) according to the manufacturer's instructions. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Finally, the designed primers (Table S2) were used for RT‐qPCR in the LightCycler 480 (Roche Diagnostics) thermal cycler, and each sample was analysed in triplicate. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | RNA‐seq was performed by BGI as described before. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | In brief, after GIST‐T1‐IR cells were treated with DMSO or β‐elemene for 12 h, the total RNA was extracted using TRIzol reagent (Invitrogen, 15596026). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | RNA‐seq and the following bioinformatic analyses were performed using the Illumina HiSeq platform (Illumina) as previously described. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Analysis and visualisation of the differentially expressed genes (DEGs) were conducted using the limma and ggplot 2 packages in R software. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was used for gene functional annotation. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | We established the criteria as the p‐value < .05 and log2 fold change > .5. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | TPP is a novel target screening method that can predict direct targets for small drug molecules. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The TPP principle uses the changes in the thermodynamic properties of protein stability that occur when ligands bind to small molecules by subjecting intact cells or protein lysates to a temperature gradient in the presence of small molecules to precipitate insoluble heat‐denatured proteins. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The soluble fraction of the protein was then recovered and quantified by LC‐mass spectrometry. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | TPP was performed according to previous reports. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The CETSA was utilised to evaluate the stability of the target protein following the previous protocol. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | GIST‐882‐IR and GIST‐T1‐IR cells were lysed by repeated freezing and thawing of the suspension in liquid nitrogen three times and centrifuging at 4°C. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The supernatant was collected and treated with Dimethyl sulfoxide(DMSO) or β‐elemene (20 µg/L) for 1 h at 25°C. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The mixtures were then divided into several tubes and heated at various temperatures ranging from 37 to 67°C for 3 min, cooled at room temperature for 3 min and then centrifuged at 4°C. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Loading buffer was added to the samples, followed by boiling at 98°C for 10 min. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The samples were subsequently analysed using Western blotting analysis. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The crystal structure of N6AMT1 used for docking was obtained from the RCSB Protein Data Bank website (https://www.rcsb.org). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | Modifications, including the elimination and hydrogenation of ethanol and water molecules, as well as the refinement of amino acids, were performed using PyMol 2.5.42. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The 3D chemical structure of β‐elemene was obtained from PubChem (https://pubchem.ncbi.nlm.nih.gov). |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | AutoDock 1.5.7 software was adopted for molecular docking. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | For docking analysis, all protein and molecular files were converted into PDBQT format. |
PMC12390768 | β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1 | The grid box was centred to cover the domain of each protein and to accommodate free molecular movement. |
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.