Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: β‐elemene induces ferroptosis in imatinib‐resistant GIST cells. (A) Chemical structure of β‐elemene. (B‐C) Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and GSEA showed that genes involved in ferroptosis were significantly dysregulated under β‐elemene treatment. (D) The Fe 2+ levels detected by FerroOrange fluorescence probe in GIST‐882‐IR and GIST‐T1‐IR cells treated with DMSO, imatinib, β‐elemene or imatinib+β‐elemene for 24 h. (E) Quantification of relative fluorescence intensity of Fe 2+ levels by Image J. (F) The MDA levels detected in GIST‐882‐IR and GIST‐T1‐IR cells treated with DMSO, imatinib, β‐elemene or imatinib+β‐elemene for 24 h. (G) Representative fluorescent images of lipid peroxidation detected by BODIPY (581/591) C11 probe in GIST‐882‐IR and GIST‐T1‐IR cells treated with DMSO, imatinib, β‐elemene or imatinib+β‐elemene for 24 h. Scale bar = 100 µm. (H) Quantification of relative fluorescence intensity of oxidised BODIPY by Image J. (I) The ROS accumulation was detected using flow cytometry analysis and statistical histograms of positive cells in GIST‐882‐IR and GIST‐T1‐IR cells treated with DMSO, imatinib, β‐elemene or imatinib+β‐elemene for 24 h. (J) Representative cell ultrastructural images of GIST‐T1‐IR cells treated with DMSO, imatinib, β‐elemene or imatinib+β‐elemene for 24 h. All experiments were performed in triplicate. Data represent the mean ± SD; * p < .05; ** p < .01; *** p < .001. An unpaired t ‐test was used unless otherwise stated.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques: Fluorescence, Flow Cytometry

Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: β‐elemene promotes imatinib sensitivity in imatinib‐resistant GISTs through ferroptosis. (A) The interaction between imatinib and β‐elemene on cell cytotoxicity was examined by the median‐effect method of Chou–Talalay. (B) The dose and combination index of imatinib in combination with β‐elemene on GIST‐882‐IR and GIST‐T1‐IR cells was estimated by calculation of combination index (CI) values using Compusyn software. The darker point indicates a stronger synergistic effect. The fraction affected values indicate the percentage of cell inhibition, while the CI values indicate the effects of combination treatments. (C) Antitumour effects of imatinib and β‐elemene in GIST‐882‐IR and GIST‐T1‐IR cells after the indicated treatment were evaluated by colony formation assay. (D) Statistical analysis of the colony formation assay. (E) Representative results of PI‐positive cells in IM‐exposed GIST‐882‐IR and GIST‐T1‐IR after the indicated treatment and quantitative analysis after the treatment for 48 h. (F) Statistical histogram of the flow cytometry cell death analysis. (G) Antitumour effects of GIST‐882‐IR and GIST‐T1‐IR cells treated with or without ferroptosis inhibitor Ferrostatin‐1 (Fer‐1) were evaluated by colony formation assay. (H) Statistical analysis of the colony formation assay of GIST‐882‐IR and GIST‐T1‐IR cells treated with or without ferroptosis inhibitor Fer‐1. (I) Cell viability of GIST‐882‐IR and GIST‐T1‐IR cells treated with or without ferroptosis inhibitor Fer‐1. (J) Schematic description of the in vivo anticancer effect of combined treatment with β‐elemene and imatinib in the cell line‐based xenograft model. (K) Photograph and comparison of tumour sizes in the indicated groups. (L) Growth curve of GIST‐T1‐IR xenografts in the indicated groups. (M) The tumour weight of GIST‐T1‐IR xenografts in the indicated groups. (N) Representative images of IHC staining of MKI67/K‐67 in mouse tumours. All experiments were performed in triplicate. Data represent the mean ± SD; * p < .05; ** p < .01; *** p < .001. An unpaired t ‐test was used unless otherwise stated.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques: Software, Inhibition, Colony Assay, Flow Cytometry, In Vivo, Comparison, Immunohistochemistry

Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: β‐elemene induced ferroptosis in imatinib‐resistant GISTs through HMOX1. (A) Heatmap of the RNA‐seq analysis results for GIST‐T1‐IR cells treated with DMSO or β‐elemene. (B) Volcano plot of down‐regulated and upregulated for GIST‐T1‐IR cells treated with DMSO or β‐elemene. (C) Western blotting analysis of HMOX1 and GPX4 expression in GIST cells treated with imatinib, β‐elemene or imatinib+β‐elemene. (D) Western blotting analysis of HMOX1 expression in parental and imatinib‐resistant GIST cells. (E) Western blotting analysis of HMOX1 in GIST‐882‐IR and GIST‐T1‐IR cells transfected with HMOX1‐expressing plasmid. (F) CCK‐8 assay of sensitivity to imatinib in HMOX1‐overexpressed GIST‐882‐IR and GIST‐T1‐IR cells versus control GIST‐882_IR and GIST‐T1‐IR cells. (G) The colony formation assay of sensitivity to imatinib in HMOX1‐overexpressed GIST‐882‐IR and GIST‐T1‐IR cells versus control GIST‐882_IR and GIST‐T1‐IR cells. (H) Western blot showing changes in HMOX1 expression in response to Hemin. (I‐J) The colony formation assay and statistical histogram of GIST‐882‐IR and GIST‐T1‐IR cells treated with imatinib+β‐elemene with or without hemin. (K) Cell death measurements by flow cytometry analysis in GIST‐882‐IR and GIST‐T1‐IR cells treated with imatinib+β‐elemene with or without hemin. (L–M) The colony formation assay and statistical histogram of GIST‐882‐IR and GIST‐T1‐IR cells treated with imatinib+β‐elemene with or without zinc protoporphyrin‐9 (ZnPP). (N) Cell death measurements by flow cytometry analysis in GIST‐882‐IR and GIST‐T1‐IR cells treated with imatinib+β‐elemene with or without ZnPP. (O) Photograph and comparison of tumour sizes in different groups. (P) Growth curve of GIST‐T1‐IR xenografts in the indicated groups. (Q) The tumour weight of GIST‐T1‐IR xenografts in the indicated groups. All experiments were performed in triplicate. Data represent the mean ± SD; * p < .05; ** p < .01; *** p < .001. An unpaired t ‐test was used unless otherwise stated.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques: RNA Sequencing, Western Blot, Expressing, Transfection, Plasmid Preparation, CCK-8 Assay, Control, Colony Assay, Flow Cytometry, Comparison

Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: β‐elemene targets N6AMT1 to promote imatinib sensitivity in imatinib‐resistant GIST cells via the nuclear factor erythroid 2‐related factor 2 (NRF2)/HMOX1 axis. (A) The workflow for cellular targets identification of β‐elemene in GIST‐T1‐IR by thermal proteome profiling (TPP). (B) Heatmap of differentially expressed protein in GIST‐T1‐IR cells between control and β‐elemene‐treated groups. (C) Venn diagram of β‐elemene target screening. (D) Volcano plot of cellular targets of β‐elemene by the TPP strategy. (E) Western blot analysis of NRF2 and HMOX1 in GIST cells with or without N6AMT1 knockdown by small interfering RNA (siRNA). (F) Western blot analysis of NRF2 and HMOX1 in GIST cells with or without N6AMT1 overexpression. (G) Cellular thermal shift assay demonstrated a stabilisation effect of N6AMT1 with β‐elemene. (H) The interaction between β‐elemene with N6AMT1 targeting Asp103 was predicted by molecular docking. (I) GIST‐T1‐IR cell lysates were incubated with either biotin or β‐elemene‐biotin at 4°C overnight, and then a pulldown assay was performed. (J) GIST‐T1‐IR cell lysates were preincubated with either DMSO or free β‐elemene, followed by subsequent incubation with β‐elemene‐biotin. The interaction between N6AMT1 and β‐elemene was then detected by capturing β‐elemene‐biotin. (K) The mutant N6AMT1 proteins were incubated with β‐elemene, followed by protein affinity pull‐down assay and detected by immunoblotting. (L) Western blot analysis of N6AMT1/NRF2/HMOX1 axis in GIST cells treated with imatinib, β‐elemene or imatinib+β‐elemene. (M) Western blot analysis of NRF2 in cytosol protein and nuclear protein. (N) The staining intensity and localisation of NRF2 in the indicated groups were analysed by immunofluorescence staining. (O) Western blot analysis of the relationship of NRF2 and HMOX1 in ferroptosis with knockdown of NRF2 in GIST cells followed by imatinib+β‐elemene treatment. (P) Methylation level of NRF2 promoter in GIST‐T1‐IR cell treated with imatinib, β‐elemene or imatinib+β‐elemene. (Q) Methylation level of NRF2 promoter in GIST‐T1‐IR cell with or without N6AMT1 knockdown by siRNA. (R) Methylation level of NRF2 promoter in GIST‐T1‐IR cell with or without N6AMT1 overexpression. Data represent the mean ± SD; * p < .05; ** p < .01; *** p < .001. An unpaired t ‐test was used unless otherwise stated.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques: Control, Western Blot, Knockdown, Small Interfering RNA, Over Expression, Thermal Shift Assay, Incubation, Mutagenesis, Pull Down Assay, Staining, Immunofluorescence, Methylation

Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: β‐elemene improves imatinib therapeutic efficiency in imatinib‐resistant GIST. (A–D) IHC staining of Nrf2, HMOX1 and 4‐HNE in tumour tissues generated by GIST‐T1‐IR cells‐based xenograft in the indicated groups. Scale bars, 50 µm. (E) Schematic description of the in vivo anticancer effect of combined treatment with imatinib and β‐elemene in the patient‐derived xenograft (PDX) model. (F) Photograph and comparison of tumour sizes of the PDX model in the indicated groups. (G) Growth curve of the PDX model in the indicated groups. (H) The tumour weight of the PDX model in the indicated groups. (I–M) IHC staining of 4‐HNE and HMOX1 in tumour tissues generated by the PDX model in the indicated groups. Scale bars, 50 µm. Data represent the mean ± SD; * p < .05; ** p < .01; *** p < .001. An unpaired t ‐test was used unless otherwise stated.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques: Immunohistochemistry, Generated, In Vivo, Derivative Assay, Comparison

Journal: Clinical and Translational Medicine

Article Title: β‐elemene promotes ferroptosis to improve the sensitivity of imatinib in gastrointestinal stromal tumours by targeting N6AMT1

doi: 10.1002/ctm2.70438

Figure Lengend Snippet: Summary diagram of the mechanism that β‐elemene increased the sensitivity of GIST cells to imatinib. β‐elemene specifically targets N6AMT1, inhibiting its transcriptional repression function and activating the NRF2‐HMOX1 signalling pathway to induce ferroptosis.

Article Snippet: All cell lines were cultured in an incubator with 37 ° C and 5% CO 2 . β‐elemene was provided by Dalian HolleyKingkong Pharmaceutical Co. Ltd. Imatinib (T6230) was purchased from TargetMol.

Techniques:

Journal: Scientific Reports

Article Title: Host plant affects the sexual attractiveness of the female white-spotted longicorn beetle, Anoplophora malasiaca

doi: 10.1038/srep29526

Figure Lengend Snippet: Total ion chromatogram of the n -hexane fractions separated from MO- ( a ) and WI-fed female ( b ) extracts. Enlarged chromatograms for each female are shown on the right. The peaks indicated with arrows were previously identified as contact sex pheromones . Peaks 1 and 2 represent β -elemene and β -caryophyllene, respectively.

Article Snippet: The candidate chemical, (−)- β -elemene (LKT Laboratories, Inc., St Paul, MN, USA; purity, 98.9%), was adjusted to 0.025, 0.1, 1, and 10 fe (1, 4, 40, and 400 ng per treatment, respectively) and applied to dummies along with the WI-fed female extract.

Techniques:

Journal: Thoracic Cancer

Article Title: FBP1 induced by β‐elemene enhances the sensitivity of gefitinib in lung cancer

doi: 10.1111/1759-7714.14750

Figure Lengend Snippet: Downregulation of FBP1 in gefitinib‐resistant lung cells. (a) Simplified β‐elemene‐key target network. (b) Heatmap depicting the differentially expressed genes between parental HCC4006 cells and gefitinib‐resistant HCC4006 cells based on GSE123066 datasheets. (c) Venn diagram of β‐elemene targets and the downregulated genes in gefitinib‐resistant HCC4006 cells from GSE123066 datasheets. (d) The mRNA and protein levels of FBP1 in PC9, PC9/GR, HCC827, and HCC827/GR cells. (e). The mRNA and protein levels of FBP1 in PC9/GR and HCC827/GR cells with or without 10 μg/ml β‐elemene treatment. (f) Representative images of the dose‐dependent increase in FBP1 expression after β‐elemene treatment. ** p < 0.01

Article Snippet: The β‐elemene injection (National Medical Product Administration Approval Number: Chinese medicine H10960114) was obtained from Dalian Huali Jingang Pharmaceutical (Dalian, China).

Techniques: Expressing

Journal: Thoracic Cancer

Article Title: FBP1 induced by β‐elemene enhances the sensitivity of gefitinib in lung cancer

doi: 10.1111/1759-7714.14750

Figure Lengend Snippet: FBP1‐mediated sensitivity to gefitinib in PC9/GR and HCC827/GR cells. PC9/GR and HCC827/GR were transduced with SCR or shFBP1 (shRNA targeting FBP1) and then treated with 10 μg/ml β‐elemene for 24 h for subsequent experiments. (a) The protein level of FBP1 was measured in PC9/GR and HCC827/GR cells. (b) Growth curve, (c) apoptosis level, (d) colony formation, and (e) IC50 values of gefitinib in PC9/GR and HCC827/GR cells as indicated treatment. ** p < 0.01, compared with PBS; # p < 0.05, compared with SCR

Article Snippet: The β‐elemene injection (National Medical Product Administration Approval Number: Chinese medicine H10960114) was obtained from Dalian Huali Jingang Pharmaceutical (Dalian, China).

Techniques: Transduction, shRNA

Journal: Thoracic Cancer

Article Title: FBP1 induced by β‐elemene enhances the sensitivity of gefitinib in lung cancer

doi: 10.1111/1759-7714.14750

Figure Lengend Snippet: β‐elemene regulates gefitinib resistance through the FBP1/STAT3 axis in PC9/GR and HCC827/GR cells. (a and b) The protein expression of STAT3 and p‐STAT3 (Tyr705) in PC9/GR (a) and HCC827/GR (b) cells after β‐elemene treatment, followed by or without 0.5 μM colivelin treatment in the nucleus. The protein expression of BCL‐2, Bax, and cleaved caspase‐3 in PC9/GR (a) and HCC827/GR (b) cells after β‐elemene treatment, followed by or without colivelin treatment in the cytoplasm. (c) Growth curve, (d) apoptosis level, (e) colony formation, and (f) IC50 values of gefitinib in PC9/GR and HCC827/GR cells as indicated treatment. ** p < 0.01, compared with PBS; # p < 0.05, compared with DMSO

Article Snippet: The β‐elemene injection (National Medical Product Administration Approval Number: Chinese medicine H10960114) was obtained from Dalian Huali Jingang Pharmaceutical (Dalian, China).

Techniques: Expressing

Journal: Thoracic Cancer

Article Title: FBP1 induced by β‐elemene enhances the sensitivity of gefitinib in lung cancer

doi: 10.1111/1759-7714.14750

Figure Lengend Snippet: The working model describes the mechanism of FBP1. The expression of FBP1 was increased in gefitinib‐resistant lung cancer cells when stimulated by β‐elemene. The upregulation of FBP1 bound to STAT3 and inhibited the phosphorylation of STAT3, resulting in the altered expression of apoptosis‐related genes and leading to gefitinib sensitivity

Article Snippet: The β‐elemene injection (National Medical Product Administration Approval Number: Chinese medicine H10960114) was obtained from Dalian Huali Jingang Pharmaceutical (Dalian, China).

Techniques: Expressing, Phospho-proteomics

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Anti-Lung-Cancer Activity and Liposome-Based Delivery Systems of β -Elemene

doi: 10.1155/2012/259523

Figure Lengend Snippet: Chemical structure of β -Elemene.

Article Snippet: Afterwards, an injection preparation containing 85% β -elemene developed by CSPC Pharmaceutical Limited Cooperation (China) has been approved by the SFDA, China, for the treatment of brain tumours and other carcinomas [ ].

Techniques:

Journal: Oncology Reports

Article Title: β-elemene inhibits the generation of peritoneum effusion in pancreatic cancer via suppression of the HIF1A-VEGFA pathway based on network pharmacology

doi: 10.3892/or.2019.7360

Figure Lengend Snippet: Network of β-elemene-target genes. Red diamond represents β-elemene; blue circles represent target genes of β-elemene.

Article Snippet: Serial concentrations of β-elemene [CSPC Pharmaceutical Group Company Ltd. (YUANDA), Dalian, China] were added to the cells: 0, 0.5, 1, 2, 4, 8 and 16 µM.

Techniques:

Journal: Oncology Reports

Article Title: β-elemene inhibits the generation of peritoneum effusion in pancreatic cancer via suppression of the HIF1A-VEGFA pathway based on network pharmacology

doi: 10.3892/or.2019.7360

Figure Lengend Snippet: Thirty-three target genes for β-elemene against pancreatic cancer. (A) Venn diagram revealing the overlapping target genes for β-elemene against pancreatic cancer. (B) Network of target genes for β-elemene against pancreatic cancer. Red diamond represents β-elemene; red hexagon represents pancreatic cancer; blue circles represent target genes of both β-elemene and pancreatic cancer; yellow triangles represent KEGG pathway analysis of 33 target genes based on the DAVID database. KEGG, Kyoto encyclopedia of genes and genomes; DAVID, Database for Annotation, Visualization and Integrated Discovery.

Article Snippet: Serial concentrations of β-elemene [CSPC Pharmaceutical Group Company Ltd. (YUANDA), Dalian, China] were added to the cells: 0, 0.5, 1, 2, 4, 8 and 16 µM.

Techniques:

Journal: Oncology Reports

Article Title: β-elemene inhibits the generation of peritoneum effusion in pancreatic cancer via suppression of the HIF1A-VEGFA pathway based on network pharmacology

doi: 10.3892/or.2019.7360

Figure Lengend Snippet: Molecular docking simulation of β-elemene to HIF1A. Sticks represent the three-dimensional structure of β-elemene; surface represents the protein structure of HIF1A. HIF1A, hypoxia-inducible factor 1-α.

Article Snippet: Serial concentrations of β-elemene [CSPC Pharmaceutical Group Company Ltd. (YUANDA), Dalian, China] were added to the cells: 0, 0.5, 1, 2, 4, 8 and 16 µM.

Techniques:

Journal: Oncology Reports

Article Title: β-elemene inhibits the generation of peritoneum effusion in pancreatic cancer via suppression of the HIF1A-VEGFA pathway based on network pharmacology

doi: 10.3892/or.2019.7360

Figure Lengend Snippet: Antitumor effect of β-elemene in pancreatic cancer. (A) MTT assay revealed that cell viability was suppressed by β-elemene in PANC-1 cells in a dose-dependent manner. (B) MTT assay revealed that cell viability was suppressed by β-elemene in BxPC3 cells in a dose-dependent manner. (C and D) MTT assay revealed that cell viability was suppressed by β-elemene in cells isolated from peritoneum effusion of pancreatic cancer patients in a dose-dependent manner.

Article Snippet: Serial concentrations of β-elemene [CSPC Pharmaceutical Group Company Ltd. (YUANDA), Dalian, China] were added to the cells: 0, 0.5, 1, 2, 4, 8 and 16 µM.

Techniques: MTT Assay, Isolation

Journal: Oncology Reports

Article Title: β-elemene inhibits the generation of peritoneum effusion in pancreatic cancer via suppression of the HIF1A-VEGFA pathway based on network pharmacology

doi: 10.3892/or.2019.7360

Figure Lengend Snippet: Validation of the HIF1A-VEGFA pathway inhibited by β-elemene in pancreatic cancer. (A) Western blot analysis in PANC-1 and BxPC3 pancreatic cancer cell lines and (B) corresponding histogram of protein expression levels of HIF1A and VEGFA in PANC-1 and BxPC3 pancreatic cancer cells. Three independent biological replicates were performed and data are presented as the mean ± standard deviation. *P<0.05 and **P<0.01. NS, not significant. HIF1A, hypoxia-inducible factor 1-α; VEGFA, vascular endothelial growth factor A.

Article Snippet: Serial concentrations of β-elemene [CSPC Pharmaceutical Group Company Ltd. (YUANDA), Dalian, China] were added to the cells: 0, 0.5, 1, 2, 4, 8 and 16 µM.

Techniques: Western Blot, Expressing, Standard Deviation