Journal: Thoracic Cancer

Article Title: Exosomal GPT2 derived from triple‐negative breast cancer cells promotes metastasis by activating BTRC

doi: 10.1111/1759-7714.14984

Figure Lengend Snippet: Isolation and identification of exosomes. (a) Transmission electron microscopy and NTA analysis of exosomes derived from BT474 and BT549. Scale bar = 100 nm. (b) Western blot analysis of exosomal positive protein marker TSG101 and glutamate pyruvate transaminase 2 (GPT2).

Article Snippet: Human breast cancer cells BT549 and BT474 were obtained from ATCC.

Techniques: Isolation, Transmission Assay, Electron Microscopy, Derivative Assay, Western Blot, Marker

Journal: Thoracic Cancer

Article Title: Exosomal GPT2 derived from triple‐negative breast cancer cells promotes metastasis by activating BTRC

doi: 10.1111/1759-7714.14984

Figure Lengend Snippet: Exosomal glutamate pyruvate transaminase 2 (GPT2) enhances the metastatic capacity of breast cancer cells. (a) Wound healing assay of BT474 and BT474‐shGPT2 treated with BT549‐exosomes (50 μg/mL) (** p < 0.01; * p < 0.05). (b) Transwell assay of BT474 and BT474‐shGPT2 treated with BT549‐exosomes (50 μg/mL) (** p < 0.01; * p < 0.05). (c) Exosomal GPT2 promoted liver metastasis of BT474 cells. Metastatic sites in the liver decreased in BT474‐shGPT2 cells treated with BT549‐exosomes (50 μg/mL) (** p < 0.01; * p < 0.05).

Article Snippet: Human breast cancer cells BT549 and BT474 were obtained from ATCC.

Techniques: Wound Healing Assay, Transwell Assay

Journal: Cells

Article Title: Inhibition of Monoacylglycerol Lipase Decreases Angiogenic Features of Endothelial Cells via Release of Tissue Inhibitor of Metalloproteinase-1 from Lung Cancer Cells.

doi: 10.3390/cells12131757

Figure Lengend Snippet: Figure 2. Effects of conditioned medium (CM) obtained from A549 cells treated with MAGL inhibitor JZL184 (A), MAGL substrate 2-AG (B), additional MAGL inhibitors JW651 (C) and MJN110 (D), or DAGL inhibitor RHC 80267 (E) on migration, viability, and tube formation of HUVECs. Migration (Boyden chamber assay, white bars), viability (WST-1 assay, black bars), and tube formation (tube formation assays, gray bars) of HUVECs were determined after incubation with CM from A549 cells for 24 h (migration and viability assay) or 2 h (tube formation analysis). The CMs used were from A549 cells previously incubated for 48 h with vehicle (Veh) or the indicated concentrations of the test compounds. All percentage values given refer to serum-free DMEM (unconditioned medium, UCM) set to 100%. Data represent mean ± SEM of n = 6 ((A,B,E), migration and tube formation), n = 3–4 (C,D), or n = 8 ((E), viability) per group. ** p ≤0.01, *** p ≤0.001 vs. UCM; # p ≤0.05, ## p ≤0.01, ### p ≤0.001 vs. CM of vehicle-treated A549 cells; one-way ANOVA with Bonferroni post hoc test.

Article Snippet: A549 human lung carcinoma cells were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany; DSMZ cat. No. ACC-107, RRID:CVCL_0023).

Techniques: Migration, Boyden Chamber Assay, WST-1 Assay, Incubation, Viability Assay

Journal: Cells

Article Title: Inhibition of Monoacylglycerol Lipase Decreases Angiogenic Features of Endothelial Cells via Release of Tissue Inhibitor of Metalloproteinase-1 from Lung Cancer Cells.

doi: 10.3390/cells12131757

Figure Lengend Snippet: Figure 3. Evaluation of the role of the endocannabinoid system in the antiangiogenic effect of JZL184 (A) and 2-AG (B) on HUVECs by testing the effect of AM-251 (CB1 antagonist), AM-630 (CB2 antagonist), and capsazepine (Capsa, TRPV1 antagonist) on the antiangiogenic effect of CM from A549 cells treated with 1 µM JZL184 (A) or 2-AG (B). A549 cells were pre-incubated with the respective receptor antagonist (all tested at a final concentration of 1 µM) for 1 h and then incubated with vehicle, JZL184, or 2-AG for 48 h before CM was collected. Migration (Boyden chamber assay, white bars), viability (WST-1 assay, black bars), and tube formation (tube formation assays, gray bars) of HUVECs were determined after incubation with CM from A549 cells for 24 h (migration and viability assay) or 2 h (tube formation analysis). The control experiment shown in (C) demonstrates the effect of CM of A549 cells previously incubated with AM-251, AM-630, and capsazepine alone for 48 h on HUVEC migration, viability, and tube formation. In the experiment shown in (D), modulation of angiogenic properties by CM of A549 cells previously treated for 48 h with vehicle, JZL184 (1 µM), palmitic acid (PA, 10 µM), or the combination of JZL184 (1 µM) and PA (10 µM) was determined. PA was used to supplement potentially reduced free fatty acids due to MAGL inhibition. All percentage values given refer to HUVECs suspended in vehicle-containing CM set to 100%. Data represent mean ± SEM of n = 9 (A), n = 3 ((B,D), tube formation), n = 6 ((C), migration, tube formation; (D), migration), n = 7 ((C), viability), or n = 4 ((D), viability) per group. * p ≤0.05, *** p ≤0.001 vs. CM of vehicle-treated A549 cells; # p ≤0.05, ## p ≤0.01, ### p ≤0.001 vs. CM of JZL184- or 2-AG-treated A549 cells; one-way ANOVA with Bonferroni (A,B,D) or Dunnett (C) post hoc test.

Article Snippet: A549 human lung carcinoma cells were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany; DSMZ cat. No. ACC-107, RRID:CVCL_0023).

Techniques: Incubation, Concentration Assay, Migration, Boyden Chamber Assay, WST-1 Assay, Viability Assay, Control, Inhibition

Journal: Cells

Article Title: Inhibition of Monoacylglycerol Lipase Decreases Angiogenic Features of Endothelial Cells via Release of Tissue Inhibitor of Metalloproteinase-1 from Lung Cancer Cells.

doi: 10.3390/cells12131757

Figure Lengend Snippet: Figure 4. Effect of JZL184 (A,B) and 2-AG (C,D) on TIMP-1 protein expression in A549 cells and angiogenic abilities of HUVECs suspended in conditioned medium (CM) of vehicle- or JZL184- treated A549 cells in the presence or absence of TIMP-1 siRNA. In (A,C), the concentration-dependent effects of JZL184 (A) and 2-AG (C) on TIMP-1 protein expression are shown. A549 cells were incubated with vehicle and the respective concentration of JZL184 or 2-AG for 48 h followed by Western blot analysis. All percentage values given refer to vehicle-treated cells set to 100%. Data (A,C) represent mean ± SEM obtained from densitometric analysis of n = 4 experiments per group. In (B,D), A549 cells were incubated with transfection reagent in the absence of any siRNA (first and second triplets or bands) and transfected with TIMP-1 siRNA (TIMP-1 si; third and fourth triplets or bands) or non-silencing siRNA (nonsi; fifth and sixth triplet or bands) for 24 h in 10% FCS containing DMEM. Thereafter, A549 cells were washed and treated with vehicle, 1 µM JZL184 (C), or 1 µM 2-AG (D) for 48 h in serum-free DMEM prior to collection of CM. Migration (Boyden chamber assay, white bars), viability (WST-1 test, black bars), and tube formation (tube formation assays, gray bars) of HUVECs was measured after suspension in CM from A549-treated cells. The incubation time of HUVECs was 24 h for migration and viability testing and 2 h for tube formation analysis. Monitoring of TIMP-1 protein was performed in parallel using CM obtained from A549 cells. The Western blot images shown here are representative of a total of three (B) or four (D) experiments performed. For analysis of angiogenesis data in (B,D), vehicle-containing CM was set at 100%. Data represent mean ± SEM of n = 3 ((B), migration, tube formation; (D)) or n = 4 ((B), viability) per group. ** p ≤0.01, *** p ≤0.001 vs. corresponding vehicle control; ## p ≤0.01, ### p ≤0.001 vs. the respective JZL184 (B) or 2-AG (D) group without siRNA; one-way ANOVA with Dunnett (A,C) or Bonferroni (B,D) post hoc test.

Article Snippet: A549 human lung carcinoma cells were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany; DSMZ cat. No. ACC-107, RRID:CVCL_0023).

Techniques: Expressing, Concentration Assay, Incubation, Western Blot, Transfection, Migration, Boyden Chamber Assay, Suspension, Control

Journal: Cells

Article Title: Inhibition of Monoacylglycerol Lipase Decreases Angiogenic Features of Endothelial Cells via Release of Tissue Inhibitor of Metalloproteinase-1 from Lung Cancer Cells.

doi: 10.3390/cells12131757

Figure Lengend Snippet: Figure 5. Effect of conditioned medium (CM) derived from another lung cancer cell line (H358) and from a non-cancerous bronchial epithelial cell line (BEAS-2B) on the angiogenic capacities of HUVECs. Migration (Boyden chamber assay, white bars), viability (WST-1 assay, black bars), and tube formation (tube formation assays, gray bars) of HUVECs were determined after suspension in CM from vehicle-, JZL184-, and 2-AG-treated H358 (A,B) or BEAS-2B cells (C,D). To generate CM, H358 or BEAS-2B cells were incubated for 48 h with vehicle (Veh) or the indicated concentrations of test compounds. The exposure time of HUVECs to the indicated CM was 24 h for the migration and viability assay and 2 h for tube formation analysis. Vehicle-containing UCM was set at 100%. Data represent mean ± SEM of n = 3 ((A–D), tube formation; (A–C), migration; (A), viability), n = 4 ((D), migration), n = 5 ((B), viability), or n = 6 ((C), viability) per group. * p ≤0.05, ** p ≤0.01, *** p ≤0.001 vs. UCM; # p ≤0.05, ## p ≤0.01, ### p ≤0.001 vs. CM of vehicle-treated A549 cells; one-way ANOVA with Bonferroni post hoc test.

Article Snippet: A549 human lung carcinoma cells were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany; DSMZ cat. No. ACC-107, RRID:CVCL_0023).

Techniques: Derivative Assay, Migration, Boyden Chamber Assay, WST-1 Assay, Suspension, Incubation, Viability Assay

Journal: Cells

Article Title: Inhibition of Monoacylglycerol Lipase Decreases Angiogenic Features of Endothelial Cells via Release of Tissue Inhibitor of Metalloproteinase-1 from Lung Cancer Cells.

doi: 10.3390/cells12131757

Figure Lengend Snippet: Figure 7. Impact of MAGL inhibitor JZL184 on the growth of A549 xenografts from nude mice as well as on the number of CD31- (angiogenesis and vascularization marker) and TIMP-1-positive cells in xenografts. Tumors were generated by subcutaneous inoculation of 1 × 107 A549 cells into the right dorsal flank. Animals were treated with either vehicle or JZL184 (4, 8, or 16 mg/kg i.p.) every 72 h for 28 days. Tumor size was measured with an external caliper and calculated as described in Materials and Methods and are shown as tumor volumes over time in panel A. The images beside the time-course were taken from representative tumors on day 28. Quantification of CD31 (B) and TIMP-1 (C) was performed by counting the positively stained cells of xenograft-derived paraffine sections stained with the indicated antibodies and calculating the percentage relative to the total number of cells per field of view. Values are means ± SEM of n = 7–8 (A), n = 4 (B), or n = 5 (C) animals per group. * p ≤0.05, ** p ≤0.01, *** p ≤0.001 vs. vehicle; one-way ANOVA with Dunnett post hoc test.

Article Snippet: A549 human lung carcinoma cells were purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany; DSMZ cat. No. ACC-107, RRID:CVCL_0023).

Techniques: Marker, Generated, Staining, Derivative Assay

Journal: The EMBO Journal

Article Title: Reciprocal priming between receptor tyrosine kinases at recycling endosomes orchestrates cellular signalling outputs

doi: 10.15252/embj.2020107182

Figure Lengend Snippet:

Article Snippet: BT549 , LGC ATCC® , HTB‐122.

Techniques: Virus, Recombinant, Extraction, Multiplex sample analysis, Transfection, Molecular Weight, Marker, Reverse Transcription, Protease Inhibitor, Membrane, Cell Culture, Clinical Proteomics, Spectroscopy, Imaging, cDNA Synthesis, Plasmid Preparation, Negative Control, Mutagenesis, Clone Assay, Generated, Software, Microscopy

Journal: Cell death & disease

Article Title: AURKAIP1 actuates tumor progression through stabilizing DDX5 in triple negative breast cancer.

doi: 10.1038/s41419-023-06115-1

Figure Lengend Snippet: Fig. 2 AURKAIP1 promoted cell proliferation and migration of triple-negative breast cancer (TNBC). A The relative mRNA and protein expression levels of AURKAIP1 in normal breast cell (MCF 10A) and five TNBC cell lines. B RT-qPCR analyses of mRNA expression level in the TNBC cells after knockdown or overexpression of AURKAIP1. C The effects induced by AURKAIP1 suppression and overexpression on colony- forming ability were detected by colony formation assays. D The CCK8 assays were performed to evaluate the cell viability of TNBC cells transfected with AURKAIP1 siRNAs or plasmids. E Images and tumor growth curves of subcutaneous tumor formation in nude mice with stable shScramble or shAURKAIP1 cells (n = 5 for each group). Data are represented with mean ± SD from three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001, ****p < 0.0001).

Article Snippet: Human TNBC cell lines (MDA-MB-231, BT 549, MDA-MB-468, MDA-MB-157, and HCC 1806), human normal breast epithelial cell line (MCF 10A) and the HEK 293T cell line were all purchased from American Tissue Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Migration, Expressing, Quantitative RT-PCR, Knockdown, Over Expression, Transfection