fao inhibitor st1326  (MedChemExpress)

Journal: Journal of Advanced Research

Article Title: Mitochondrial fatty acid oxidation as the target for blocking therapy-resistance and inhibiting tumor recurrence: The proof-of-principle model demonstrated for ovarian cancer cells

doi: 10.1016/j.jare.2025.03.026

Figure Lengend Snippet: Enhanced FAO in paclitaxel-tolerant ovarian cancer persister cells. ( A ) Bubble diagram of the altered proteins between OVCAR8 and OVCAR8-Persister cells in KEGG analysis. ( B ) GSEA of the fatty acid beta-oxidation (GO: 0006635) gene set in OVCAR8-Persister cells compared with OVCAR8 cells. ( C and D ) Relative NADPH/NADP + ratio (C) and free fatty acid levels (D) were measured in parental and persister cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or ST1326 (5, 10 or 20 μM) treatment for 72 h. ( E ) Time series of OCR measurements in parental and persister cells derived from OVCAR8 (left) and A2780 (right) cell lines in the presence of vehicle or the FAO inhibitor ST1326, as determined via a Seahorse analyzer. ( F ) Quantification of basal respiration, maximal respiration and spare respiration capacity based on the OCR measurements. ( G ) Quantification of decreased basal respiration, maximal respiration and spare respiration capacity after the addition of ST1326. ( H ) Parental and persister cells were treated with vehicle or paclitaxel (20 nM) for 24 h. After incubation with MitoSOX™ Red (500 nM) for 30 min, representative histogram (left) and quantification (right) of MitoSOX™ Red fluorescence were performed via flow cytometry. ( I ) OVCAR8-Persister (left) and A2780-Persister (right) cells were pretreated with vehicle, ST1326 (20 μM), ERG240 (60 μM) or IACS-010759 (2 nM) for 24 h and subsequently treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay or CellTiter-Glo® assay (IACS-010759-treated group). KEGG, Kyoto Encyclopedia of Genes and Genomes; FDR, false discovery rate; ES, enrichment score; NES, normalized enrichment score; ATP, adenosine triphosphate; OCR, oxygen consumption rate; Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; Rot/AA, rotenone/antimycin A; PTX, paclitaxel. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

Article Snippet: The FAO inhibitor ST1326 (Teglicar, Cat. HY-16482, MedChemExpress, USA) was prepared as a 20 mM stock solution in double distilled H 2 O (ddH 2 O); Trimetazidine (TMZ, Cat. HY-B0968A, MedChemExpress, USA) was prepared as a 200 mM stock solution in DMSO.

Techniques: Derivative Assay, Incubation, Fluorescence, Flow Cytometry, CCK-8 Assay, Glo Assay

Journal: Journal of Advanced Research

Article Title: Mitochondrial fatty acid oxidation as the target for blocking therapy-resistance and inhibiting tumor recurrence: The proof-of-principle model demonstrated for ovarian cancer cells

doi: 10.1016/j.jare.2025.03.026

Figure Lengend Snippet: Persister cells were vulnerable to the FAO inhibitor ST1326 both in vitro and in vivo . ( A and B ) Persister cells were preexposed to vehicle or ST1326 (20 μM) for 24 h and subsequently treated with paclitaxel (20 nM) for 24 h. After incubation with MitoSOX™ Red (500 nM) for 30 min, representative images (A) were acquired with a fluorescence microscope (100 × ), and representative histogram (left) and quantification (right) of MitoSOX™ Red fluorescence (B) were performed via flow cytometry. Scale bar, 100 μm. ( C and D ) OVCAR8-Persister (C) and A2780-Persister (D) cells were pretreated with vehicle or ST1326 (20 μM) for 24 h and subsequently treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, after which cell viability was assessed using a CCK-8 assay. ( E ) Representative images (left) and quantification (right) of colonies of persister cells pretreated with vehicle or ST1326 for 24 h and subsequently treated with vehicle or paclitaxel for 48 h. ( F ) Quantification of annexin V-positive apoptotic OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle, paclitaxel or ST1326 treatment. ( G ) Western blot analysis of PARP, caspase-3, Bcl-2, BAX and cleaved-caspase-3 in OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle, paclitaxel or ST1326 treatment. ( H ) Schematic representation showing the design of the mouse experiment. NOD/SCID mice underwent orthotopic implantation of OVCAR8-luc cells and ovariectomy, followed by drug treatment twice per week for 3 weeks. Tumor growth was monitored weekly using bioluminescence imaging. ( I ) Representative bioluminescence images of the tumor burden in NOD/SCID mice at week 5 for the control and ST1326 groups, and at week 7 for the PTX and PTX + ST1326 groups. ( J ) Quantitative analysis of total photon flux in OVCAR8-luc cell xenograft-bearing mice under the indicated treatments. ( K ) Weight curves of the mice in the 4 groups. ( L ) HE-stained images (400 × ) of major organs (heart, liver, spleen, lung and kidney) from the mice in the 4 groups. Scale bar, 50 μm. β-Actin served as the internal control for the western blots. PTX, paclitaxel. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

Article Snippet: The FAO inhibitor ST1326 (Teglicar, Cat. HY-16482, MedChemExpress, USA) was prepared as a 20 mM stock solution in double distilled H 2 O (ddH 2 O); Trimetazidine (TMZ, Cat. HY-B0968A, MedChemExpress, USA) was prepared as a 200 mM stock solution in DMSO.

Techniques: In Vitro, In Vivo, Incubation, Fluorescence, Microscopy, Flow Cytometry, CCK-8 Assay, Western Blot, Imaging, Control, Staining