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ovarian adenocarcinoma cell line ovcar8  (ATCC)


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    ATCC ovarian adenocarcinoma cell line ovcar8
    Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) <t>OVCAR8</t> and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.
    Ovarian Adenocarcinoma Cell Line Ovcar8, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ovarian adenocarcinoma cell line ovcar8/product/ATCC
    Average 97 stars, based on 1149 article reviews
    ovarian adenocarcinoma cell line ovcar8 - by Bioz Stars, 2026-02
    97/100 stars

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    1) Product Images from "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"

    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

    Journal: Journal of Advanced Research

    doi: 10.1016/j.jare.2025.03.026

    Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.
    Figure Legend Snippet: Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Techniques Used: Microscopy, Alamar Blue Assay, Dilution Assay, Fluorescence, Derivative Assay, CCK-8 Assay, Concentration Assay

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    HADHA upregulation enhanced FAO and decreased paclitaxel sensitivity in persister cells. ( A ) Heatmap of proteins differentially regulated between OVCAR8 and OVCAR8-Persister cells involved in the FAO process. Red, upregulated; green, downregulated. ( B ) RT–qPCR of FAO-related genes in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( C ) Western blot analysis of HADHA and CPT1A in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( D and E ) RT–qPCR (D) and western blot analysis (E) of HADHA in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( F ) RT–qPCR and western blot analysis of HADHA in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( G ) RT–qPCR and western blot analysis of HADHA in pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( H ) Time series of OCR measurements in OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 determined via a Seahorse analyzer. ( I ) Quantification of basal respiration, maximal respiration and spare respiration capacity based on the OCR measurements. ( J ) Representative images (100 × ) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated OVCAR8-Persister (top) and A2780-Persister (bottom) cells. Scale bar, 100 μm. ( K ) Representative histograms (left) and quantification (right) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated persister cells. ( L ) OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using a CCK-8 assay. ( M ) quantification of colonies of pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells treated with vehicle or paclitaxel for 48 h. ( N ) Quantification of annexin V-positive apoptotic cells in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. ( O ) Western blot analysis of PARP, caspase-3, Bcl-2, BAX and cleaved caspase-3 in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. β-Actin served as the internal control for the RT–qPCR and western blots. NC, negative control; PTX, paclitaxel; OCR, oxygen consumption rate; Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; Rot/AA, rotenone/antimycin A. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.
    Figure Legend Snippet: HADHA upregulation enhanced FAO and decreased paclitaxel sensitivity in persister cells. ( A ) Heatmap of proteins differentially regulated between OVCAR8 and OVCAR8-Persister cells involved in the FAO process. Red, upregulated; green, downregulated. ( B ) RT–qPCR of FAO-related genes in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( C ) Western blot analysis of HADHA and CPT1A in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( D and E ) RT–qPCR (D) and western blot analysis (E) of HADHA in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( F ) RT–qPCR and western blot analysis of HADHA in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( G ) RT–qPCR and western blot analysis of HADHA in pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( H ) Time series of OCR measurements in OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 determined via a Seahorse analyzer. ( I ) Quantification of basal respiration, maximal respiration and spare respiration capacity based on the OCR measurements. ( J ) Representative images (100 × ) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated OVCAR8-Persister (top) and A2780-Persister (bottom) cells. Scale bar, 100 μm. ( K ) Representative histograms (left) and quantification (right) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated persister cells. ( L ) OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using a CCK-8 assay. ( M ) quantification of colonies of pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells treated with vehicle or paclitaxel for 48 h. ( N ) Quantification of annexin V-positive apoptotic cells in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. ( O ) Western blot analysis of PARP, caspase-3, Bcl-2, BAX and cleaved caspase-3 in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. β-Actin served as the internal control for the RT–qPCR and western blots. NC, negative control; PTX, paclitaxel; OCR, oxygen consumption rate; Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; Rot/AA, rotenone/antimycin A. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Techniques Used: Quantitative RT-PCR, Derivative Assay, Western Blot, Transfection, Stable Transfection, Fluorescence, CCK-8 Assay, Control, Negative Control

    CEBPB promoted HADHA transcription in persister cells. ( A ) Venn diagram showing the shared predicted transcription factors of HADHA according to the JASPAR, AnimalTFDB and hTFtarget databases. ( B ) Scatter plots of CEBPB vs. HADHA expression in tumor specimens from HGSOC patients treated with paclitaxel from the TCGA database. The P value and Pearson correlation coefficient (R) were shown. ( C ) Western blot analysis of CEBPB in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( D and E ) Relative expression of CEBPB and HADHA in OVCAR8-Persister (left) and A2780-Persister (right) cells 48 h after si-NC or si-C1/si-C2 transfection determined via RT–qPCR (D) and western blots (E). ( F and G ) RT–qPCR (F) and western blot analysis (G) of HADHA and Flag-CEBPB in EV- or Flag-tagged pcDNA3.1 (+)-CEBPB-transfected OVCAR8 (left) and A2780 (right) cells. ( H ) Relative expression of CEBPB and HADHA in two HGSOC PDOs 48 h after si-NC or si-C1/si-C2 transfection. ( I ) Luciferase reporter assays were conducted by cotransfecting the pGL4.21-HADHA promoter luciferase reporter with si-NC or si-C1/si-C2, as well as a Renilla luciferase reporter, into OVCAR8-Persister and A2780-Persister cells. ( J ) Schematic diagrams of deletion constructs spanning the −2,000 – +500 region of the HADHA promoter. ( K ) The pGL4.21 vector containing full-length or sequential deletion of the HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which luciferase activity was detected. ( L ) The luciferase vector pGL4.21 containing the wild-type or mutant (MUT) HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which the luciferase activity was detected. ( M ) ChIP-qPCR (left) and ChIP-PCR (right) analysis of CEBPB binding to HADHA promoter using a primer pair for the CEBPB-binding site 3. Aliquots of each PCR product were electrophoresed on a 2 % agarose gel. ( N ) RT–qPCR (top) and western blot analysis (bottom) of Flag-HADHA in EV- or pCMV-Flag-HADHA-transfected OVCAR8 (left) and A2780 (right) cells. ( O ) OVCAR8-Persister (left) and A2780-Persister (right) cells cotransfected with si-NC or si-C1/si-C2 together with EV or pCMV-Flag-HADHA vector were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. ( P ) OVCAR8 (left) and A2780 (right) cells cotransfected with EV or Flag-tagged pcDNA3.1 (+)-CEBPB vector together with si-NC or si-H1/si-H2 were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. β-Actin served as the internal control for the RT–qPCR and western blots. PDO, patient-derived organoid; NC, negative control; EV, empty vector; TSS, transcription start site; MUT, mutant. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.
    Figure Legend Snippet: CEBPB promoted HADHA transcription in persister cells. ( A ) Venn diagram showing the shared predicted transcription factors of HADHA according to the JASPAR, AnimalTFDB and hTFtarget databases. ( B ) Scatter plots of CEBPB vs. HADHA expression in tumor specimens from HGSOC patients treated with paclitaxel from the TCGA database. The P value and Pearson correlation coefficient (R) were shown. ( C ) Western blot analysis of CEBPB in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( D and E ) Relative expression of CEBPB and HADHA in OVCAR8-Persister (left) and A2780-Persister (right) cells 48 h after si-NC or si-C1/si-C2 transfection determined via RT–qPCR (D) and western blots (E). ( F and G ) RT–qPCR (F) and western blot analysis (G) of HADHA and Flag-CEBPB in EV- or Flag-tagged pcDNA3.1 (+)-CEBPB-transfected OVCAR8 (left) and A2780 (right) cells. ( H ) Relative expression of CEBPB and HADHA in two HGSOC PDOs 48 h after si-NC or si-C1/si-C2 transfection. ( I ) Luciferase reporter assays were conducted by cotransfecting the pGL4.21-HADHA promoter luciferase reporter with si-NC or si-C1/si-C2, as well as a Renilla luciferase reporter, into OVCAR8-Persister and A2780-Persister cells. ( J ) Schematic diagrams of deletion constructs spanning the −2,000 – +500 region of the HADHA promoter. ( K ) The pGL4.21 vector containing full-length or sequential deletion of the HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which luciferase activity was detected. ( L ) The luciferase vector pGL4.21 containing the wild-type or mutant (MUT) HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which the luciferase activity was detected. ( M ) ChIP-qPCR (left) and ChIP-PCR (right) analysis of CEBPB binding to HADHA promoter using a primer pair for the CEBPB-binding site 3. Aliquots of each PCR product were electrophoresed on a 2 % agarose gel. ( N ) RT–qPCR (top) and western blot analysis (bottom) of Flag-HADHA in EV- or pCMV-Flag-HADHA-transfected OVCAR8 (left) and A2780 (right) cells. ( O ) OVCAR8-Persister (left) and A2780-Persister (right) cells cotransfected with si-NC or si-C1/si-C2 together with EV or pCMV-Flag-HADHA vector were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. ( P ) OVCAR8 (left) and A2780 (right) cells cotransfected with EV or Flag-tagged pcDNA3.1 (+)-CEBPB vector together with si-NC or si-H1/si-H2 were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. β-Actin served as the internal control for the RT–qPCR and western blots. PDO, patient-derived organoid; NC, negative control; EV, empty vector; TSS, transcription start site; MUT, mutant. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Techniques Used: Expressing, Western Blot, Transfection, Quantitative RT-PCR, Luciferase, Construct, Plasmid Preparation, Activity Assay, Mutagenesis, ChIP-qPCR, Binding Assay, Agarose Gel Electrophoresis, CCK-8 Assay, Control, Derivative Assay, Negative Control



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    ovarian adenocarcinoma cell line ovcar8  (ATCC)
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    Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) <t>OVCAR8</t> and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.
    Ovarian Adenocarcinoma Cell Line Ovcar8, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    CLS Cell Lines Service GmbH ovcar8
    a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.
    Ovcar8, supplied by CLS Cell Lines Service GmbH, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ovcar8 dr sidong huang  (ATCC)
    99
    ATCC ovcar8 dr sidong huang
    CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), <t>OVCAR8</t> ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.
    Ovcar8 Dr Sidong Huang, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nih ovcar8  (ATCC)
    99
    ATCC nih ovcar8
    CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), <t>OVCAR8</t> ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.
    Nih Ovcar8, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ovarian cancer cell line ovcar8 nih n a human ovarian cancer cell line skov3 atcc htb  (ATCC)
    99
    ATCC ovarian cancer cell line ovcar8 nih n a human ovarian cancer cell line skov3 atcc htb
    CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), <t>OVCAR8</t> ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.
    Ovarian Cancer Cell Line Ovcar8 Nih N A Human Ovarian Cancer Cell Line Skov3 Atcc Htb, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cell lines ovcar8 atcc  (ATCC)
    93
    ATCC cell lines ovcar8 atcc
    CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), <t>OVCAR8</t> ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.
    Cell Lines Ovcar8 Atcc, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ovcar8  (Creative Biolabs)
    90
    Creative Biolabs ovcar8
    ( A ) Gene and pathways enrichment analysis of 1Ai-3638 and 1Ai-8214 downregulated genes by the Gene Analytics tool. ( B ) <t>OVCAR8</t> ovarian carcinoma cell line was incubated with increasing concentrations of 1Ais for 72 h, and then cell viability was assessed. Viability (100%) is normalized to 0 concentration (DMSO), n = 3 independent biological replicates, and error bars represent SD. ( C ) 1Ais toxicity assessment. Mice weight was measured at different time points after 1Ai-3638 and 1Ai-8214 subcutaneous injection for 3 weeks (assessed every 3–4 days, 3 mice per concentration, 21 mice in total) ( D ) OVCAR8 cell-derived xenografts weight after 1Ais treatment (10 mice per group). OVCAR8 cells were injected into the ovaries of immunocompromised female mice. Out of ten mice in each group (1Ai-3638,1Ai-8214 and DMSO) nine were included as they presented an ovarian tumor. 1Ais were injected into the mice’s peritoneal cavity five times a week for 4 weeks. Finally, mice were sacrificed, and tumor weight was measured. Results are presented as boxplots in which the center = median, box ends = interquartile range (IQR), bottom whisker = minimum and upper whisker= maximum, n = 9. Weight monitoring of mice during the experiment is presented in Appendix Fig. S . * P < 0.05, ** P < 0.01, Student t test. .
    Ovcar8, supplied by Creative Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    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: Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Article Snippet: The human ovarian adenocarcinoma cell line OVCAR8 (RRID:CVCL_1629) and the human embryonic kidney cell line HEK-293 T (RRID:CVCL_0063) were purchased from American Type Culture Collection (ATCC).

    Techniques: Microscopy, Alamar Blue Assay, Dilution Assay, Fluorescence, Derivative Assay, CCK-8 Assay, Concentration Assay

    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.

    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 human ovarian adenocarcinoma cell line OVCAR8 (RRID:CVCL_1629) and the human embryonic kidney cell line HEK-293 T (RRID:CVCL_0063) were purchased from American Type Culture Collection (ATCC).

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

    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.

    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 human ovarian adenocarcinoma cell line OVCAR8 (RRID:CVCL_1629) and the human embryonic kidney cell line HEK-293 T (RRID:CVCL_0063) were purchased from American Type Culture Collection (ATCC).

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

    HADHA upregulation enhanced FAO and decreased paclitaxel sensitivity in persister cells. ( A ) Heatmap of proteins differentially regulated between OVCAR8 and OVCAR8-Persister cells involved in the FAO process. Red, upregulated; green, downregulated. ( B ) RT–qPCR of FAO-related genes in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( C ) Western blot analysis of HADHA and CPT1A in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( D and E ) RT–qPCR (D) and western blot analysis (E) of HADHA in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( F ) RT–qPCR and western blot analysis of HADHA in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( G ) RT–qPCR and western blot analysis of HADHA in pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( H ) Time series of OCR measurements in OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 determined via a Seahorse analyzer. ( I ) Quantification of basal respiration, maximal respiration and spare respiration capacity based on the OCR measurements. ( J ) Representative images (100 × ) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated OVCAR8-Persister (top) and A2780-Persister (bottom) cells. Scale bar, 100 μm. ( K ) Representative histograms (left) and quantification (right) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated persister cells. ( L ) OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using a CCK-8 assay. ( M ) quantification of colonies of pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells treated with vehicle or paclitaxel for 48 h. ( N ) Quantification of annexin V-positive apoptotic cells in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. ( O ) Western blot analysis of PARP, caspase-3, Bcl-2, BAX and cleaved caspase-3 in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. β-Actin served as the internal control for the RT–qPCR and western blots. NC, negative control; PTX, paclitaxel; OCR, oxygen consumption rate; Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; Rot/AA, rotenone/antimycin A. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    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: HADHA upregulation enhanced FAO and decreased paclitaxel sensitivity in persister cells. ( A ) Heatmap of proteins differentially regulated between OVCAR8 and OVCAR8-Persister cells involved in the FAO process. Red, upregulated; green, downregulated. ( B ) RT–qPCR of FAO-related genes in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( C ) Western blot analysis of HADHA and CPT1A in parental and persister cells derived from OVCAR8 (top) and A2780 (bottom) cell lines. ( D and E ) RT–qPCR (D) and western blot analysis (E) of HADHA in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( F ) RT–qPCR and western blot analysis of HADHA in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( G ) RT–qPCR and western blot analysis of HADHA in pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells. ( H ) Time series of OCR measurements in OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 determined via a Seahorse analyzer. ( I ) Quantification of basal respiration, maximal respiration and spare respiration capacity based on the OCR measurements. ( J ) Representative images (100 × ) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated OVCAR8-Persister (top) and A2780-Persister (bottom) cells. Scale bar, 100 μm. ( K ) Representative histograms (left) and quantification (right) of MitoSOX™ Red fluorescence in si-NC- or si-H1/si-H2-transfected and paclitaxel-treated persister cells. ( L ) OVCAR8-Persister (left) and A2780-Persister (right) cells transfected with si-NC or si-H1/si-H2 were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using a CCK-8 assay. ( M ) quantification of colonies of pLKO.1-sh-NC or pLKO.1-sh-H1/sh-H2 stably transfected OVCAR8-Persister (left) and A2780-Persister (right) cells treated with vehicle or paclitaxel for 48 h. ( N ) Quantification of annexin V-positive apoptotic cells in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. ( O ) Western blot analysis of PARP, caspase-3, Bcl-2, BAX and cleaved caspase-3 in si-NC- or si-H1/si-H2-transfected OVCAR8-Persister (left) and A2780-Persister (right) cells after vehicle or paclitaxel treatment for 48 h. β-Actin served as the internal control for the RT–qPCR and western blots. NC, negative control; PTX, paclitaxel; OCR, oxygen consumption rate; Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; Rot/AA, rotenone/antimycin A. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Article Snippet: The human ovarian adenocarcinoma cell line OVCAR8 (RRID:CVCL_1629) and the human embryonic kidney cell line HEK-293 T (RRID:CVCL_0063) were purchased from American Type Culture Collection (ATCC).

    Techniques: Quantitative RT-PCR, Derivative Assay, Western Blot, Transfection, Stable Transfection, Fluorescence, CCK-8 Assay, Control, Negative Control

    CEBPB promoted HADHA transcription in persister cells. ( A ) Venn diagram showing the shared predicted transcription factors of HADHA according to the JASPAR, AnimalTFDB and hTFtarget databases. ( B ) Scatter plots of CEBPB vs. HADHA expression in tumor specimens from HGSOC patients treated with paclitaxel from the TCGA database. The P value and Pearson correlation coefficient (R) were shown. ( C ) Western blot analysis of CEBPB in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( D and E ) Relative expression of CEBPB and HADHA in OVCAR8-Persister (left) and A2780-Persister (right) cells 48 h after si-NC or si-C1/si-C2 transfection determined via RT–qPCR (D) and western blots (E). ( F and G ) RT–qPCR (F) and western blot analysis (G) of HADHA and Flag-CEBPB in EV- or Flag-tagged pcDNA3.1 (+)-CEBPB-transfected OVCAR8 (left) and A2780 (right) cells. ( H ) Relative expression of CEBPB and HADHA in two HGSOC PDOs 48 h after si-NC or si-C1/si-C2 transfection. ( I ) Luciferase reporter assays were conducted by cotransfecting the pGL4.21-HADHA promoter luciferase reporter with si-NC or si-C1/si-C2, as well as a Renilla luciferase reporter, into OVCAR8-Persister and A2780-Persister cells. ( J ) Schematic diagrams of deletion constructs spanning the −2,000 – +500 region of the HADHA promoter. ( K ) The pGL4.21 vector containing full-length or sequential deletion of the HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which luciferase activity was detected. ( L ) The luciferase vector pGL4.21 containing the wild-type or mutant (MUT) HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which the luciferase activity was detected. ( M ) ChIP-qPCR (left) and ChIP-PCR (right) analysis of CEBPB binding to HADHA promoter using a primer pair for the CEBPB-binding site 3. Aliquots of each PCR product were electrophoresed on a 2 % agarose gel. ( N ) RT–qPCR (top) and western blot analysis (bottom) of Flag-HADHA in EV- or pCMV-Flag-HADHA-transfected OVCAR8 (left) and A2780 (right) cells. ( O ) OVCAR8-Persister (left) and A2780-Persister (right) cells cotransfected with si-NC or si-C1/si-C2 together with EV or pCMV-Flag-HADHA vector were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. ( P ) OVCAR8 (left) and A2780 (right) cells cotransfected with EV or Flag-tagged pcDNA3.1 (+)-CEBPB vector together with si-NC or si-H1/si-H2 were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. β-Actin served as the internal control for the RT–qPCR and western blots. PDO, patient-derived organoid; NC, negative control; EV, empty vector; TSS, transcription start site; MUT, mutant. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    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: CEBPB promoted HADHA transcription in persister cells. ( A ) Venn diagram showing the shared predicted transcription factors of HADHA according to the JASPAR, AnimalTFDB and hTFtarget databases. ( B ) Scatter plots of CEBPB vs. HADHA expression in tumor specimens from HGSOC patients treated with paclitaxel from the TCGA database. The P value and Pearson correlation coefficient (R) were shown. ( C ) Western blot analysis of CEBPB in both parental and persister cells treated with paclitaxel for 0, 24 or 48 h. ( D and E ) Relative expression of CEBPB and HADHA in OVCAR8-Persister (left) and A2780-Persister (right) cells 48 h after si-NC or si-C1/si-C2 transfection determined via RT–qPCR (D) and western blots (E). ( F and G ) RT–qPCR (F) and western blot analysis (G) of HADHA and Flag-CEBPB in EV- or Flag-tagged pcDNA3.1 (+)-CEBPB-transfected OVCAR8 (left) and A2780 (right) cells. ( H ) Relative expression of CEBPB and HADHA in two HGSOC PDOs 48 h after si-NC or si-C1/si-C2 transfection. ( I ) Luciferase reporter assays were conducted by cotransfecting the pGL4.21-HADHA promoter luciferase reporter with si-NC or si-C1/si-C2, as well as a Renilla luciferase reporter, into OVCAR8-Persister and A2780-Persister cells. ( J ) Schematic diagrams of deletion constructs spanning the −2,000 – +500 region of the HADHA promoter. ( K ) The pGL4.21 vector containing full-length or sequential deletion of the HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which luciferase activity was detected. ( L ) The luciferase vector pGL4.21 containing the wild-type or mutant (MUT) HADHA promoter was transfected into OVCAR8 (left) and A2780 (right) cells, after which the luciferase activity was detected. ( M ) ChIP-qPCR (left) and ChIP-PCR (right) analysis of CEBPB binding to HADHA promoter using a primer pair for the CEBPB-binding site 3. Aliquots of each PCR product were electrophoresed on a 2 % agarose gel. ( N ) RT–qPCR (top) and western blot analysis (bottom) of Flag-HADHA in EV- or pCMV-Flag-HADHA-transfected OVCAR8 (left) and A2780 (right) cells. ( O ) OVCAR8-Persister (left) and A2780-Persister (right) cells cotransfected with si-NC or si-C1/si-C2 together with EV or pCMV-Flag-HADHA vector were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. ( P ) OVCAR8 (left) and A2780 (right) cells cotransfected with EV or Flag-tagged pcDNA3.1 (+)-CEBPB vector together with si-NC or si-H1/si-H2 were treated with paclitaxel for 48 h, and cell viability was assessed using a CCK-8 assay. β-Actin served as the internal control for the RT–qPCR and western blots. PDO, patient-derived organoid; NC, negative control; EV, empty vector; TSS, transcription start site; MUT, mutant. ns, not statistically significant; * P < 0.05; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

    Article Snippet: The human ovarian adenocarcinoma cell line OVCAR8 (RRID:CVCL_1629) and the human embryonic kidney cell line HEK-293 T (RRID:CVCL_0063) were purchased from American Type Culture Collection (ATCC).

    Techniques: Expressing, Western Blot, Transfection, Quantitative RT-PCR, Luciferase, Construct, Plasmid Preparation, Activity Assay, Mutagenesis, ChIP-qPCR, Binding Assay, Agarose Gel Electrophoresis, CCK-8 Assay, Control, Derivative Assay, Negative Control

    a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and mRFP-OVCAR8 nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

    doi: 10.1038/s41467-025-62296-7

    Figure Lengend Snippet: a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and mRFP-OVCAR8 nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.

    Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

    Techniques: Staining, Comparison

    a pFAK, pMLC, YAP/TAZ (green; left to right) and nuclei (blue) staining of OVCAR8 cells in collagen or organo-hydrogel (OHG) after 48 h in culture (representative images from n = 3 gels). b Quantification of immunofluorescence signal intensity of cytoplasmic pFAK, pMLC, relative to the average in collagen and cytoplasmic:nuclear ratio of YAP/TAZ in OVCAR8 cells embedded in collagen or OHG for 48 h ( n = 24 cells). c F-actin (yellow), nucleus (blue) and BODIPY (grey) staining of an OVCAR8 cell spread at the ECM-microdroplet interface. d Shape descriptors of OVCAR8 nuclei after 24 h in collagen or OHG ( n = 53 nuclei). e Relative spheroid area of OVCAR8 after 7 d in OHG ( n = 10 spheroids). f BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells (49 μm into the tissue from the surface) after 7 d in culture (representative images from n = 3 experiments). g Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. h Nuclei (red) of OVCAR8 cells in OHGs after 7 d treatment (representative images from n = 14 spheroids). i Quantification of spheroid area relative to DMSO control of OVCAR8 cells in OHGs ( n = 14 spheroids). i F-actin (green) and nuclei staining of CAOV3 spheroids embedded in OHGs for 7 d (representative images from n = 6 spheroids). k Quantification of spheroid area in CAOV3 cells in OHGs after 7 d ( n = 6 spheroids). l F-actin (green) and nuclei (blue) staining of Kuramochi spheroids in collagen or OHG for 7 d with or without TGFβ (representative images from n = 3 spheroids). For the data in ( b , d , g , i and k ), a two-sided unpaired t test was performed. For data in ( e ), a one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed. Scale bars, 25 μm ( a , c ), 100 μm ( f , I ), 200 μm ( h , j ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

    doi: 10.1038/s41467-025-62296-7

    Figure Lengend Snippet: a pFAK, pMLC, YAP/TAZ (green; left to right) and nuclei (blue) staining of OVCAR8 cells in collagen or organo-hydrogel (OHG) after 48 h in culture (representative images from n = 3 gels). b Quantification of immunofluorescence signal intensity of cytoplasmic pFAK, pMLC, relative to the average in collagen and cytoplasmic:nuclear ratio of YAP/TAZ in OVCAR8 cells embedded in collagen or OHG for 48 h ( n = 24 cells). c F-actin (yellow), nucleus (blue) and BODIPY (grey) staining of an OVCAR8 cell spread at the ECM-microdroplet interface. d Shape descriptors of OVCAR8 nuclei after 24 h in collagen or OHG ( n = 53 nuclei). e Relative spheroid area of OVCAR8 after 7 d in OHG ( n = 10 spheroids). f BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells (49 μm into the tissue from the surface) after 7 d in culture (representative images from n = 3 experiments). g Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. h Nuclei (red) of OVCAR8 cells in OHGs after 7 d treatment (representative images from n = 14 spheroids). i Quantification of spheroid area relative to DMSO control of OVCAR8 cells in OHGs ( n = 14 spheroids). i F-actin (green) and nuclei staining of CAOV3 spheroids embedded in OHGs for 7 d (representative images from n = 6 spheroids). k Quantification of spheroid area in CAOV3 cells in OHGs after 7 d ( n = 6 spheroids). l F-actin (green) and nuclei (blue) staining of Kuramochi spheroids in collagen or OHG for 7 d with or without TGFβ (representative images from n = 3 spheroids). For the data in ( b , d , g , i and k ), a two-sided unpaired t test was performed. For data in ( e ), a one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed. Scale bars, 25 μm ( a , c ), 100 μm ( f , I ), 200 μm ( h , j ). Source data are provided as a Source Data file.

    Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

    Techniques: Staining, Immunofluorescence, Control

    a Schematic representation of norbornene-functionalised hyaluronic acid (NB-HA)-based organohydrogels (OHGs) presenting matrix metalloproteinase (MMP)-degradable crosslinking peptides and cell adhesion ligands. b Storage moduli of collagen- and NB-HA-based OHGs ( n = 5 collagen-based and n = 3 NB-HA-based OHGs; average ± s.e.m.). c Normalised stress-relaxation curves of collagen- and HA-NB OHG ( n = 5 collagen-based and n = 3 NB-HA-based OHGs). d F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen- or NB-HA-based OHGs for 7 d (representative images from n = 9 spheroids). e OVCAR8 spheroid area in collagen- or NB-HA-based OHGs relative to collagen-based OHG average ( n = 9 spheroids). f F-actin (grey) and nuclei (red) of OVCAR8 cells embedded in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). g BODIPY (green), nuclei (red), and F-actin (grey) of OVCAR8 in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). Arrowheads indicate cells in direct contact with oil microdroplets. h OVCAR8 spheroid area in GFOGER- or RGD-presenting NB-HA OHG after 7 d culture relative to GFOGER OHG average ( n = 9 spheroids). i F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen-based OHGs for 7 d (representative images from n = 8 spheroids). j OVCAR8 spheroid area in collagen-based OHGs after 7 d culture relative to DMSO control ( n = 8 spheroids). k BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion depth into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m OVCAR8 spheroid area quantifications in MMP-cleavable or non-cleavable NB-HA OHG after 7 d culture relative to MMP-cleavable control ( n = 11 spheroids). For the data in ( b , e , h , j and l ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( m ). Error bars in ( b ) represent the s.e.m. Scale bars, 100 μm ( g , k ), 200 μm ( d , f , i ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

    doi: 10.1038/s41467-025-62296-7

    Figure Lengend Snippet: a Schematic representation of norbornene-functionalised hyaluronic acid (NB-HA)-based organohydrogels (OHGs) presenting matrix metalloproteinase (MMP)-degradable crosslinking peptides and cell adhesion ligands. b Storage moduli of collagen- and NB-HA-based OHGs ( n = 5 collagen-based and n = 3 NB-HA-based OHGs; average ± s.e.m.). c Normalised stress-relaxation curves of collagen- and HA-NB OHG ( n = 5 collagen-based and n = 3 NB-HA-based OHGs). d F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen- or NB-HA-based OHGs for 7 d (representative images from n = 9 spheroids). e OVCAR8 spheroid area in collagen- or NB-HA-based OHGs relative to collagen-based OHG average ( n = 9 spheroids). f F-actin (grey) and nuclei (red) of OVCAR8 cells embedded in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). g BODIPY (green), nuclei (red), and F-actin (grey) of OVCAR8 in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). Arrowheads indicate cells in direct contact with oil microdroplets. h OVCAR8 spheroid area in GFOGER- or RGD-presenting NB-HA OHG after 7 d culture relative to GFOGER OHG average ( n = 9 spheroids). i F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen-based OHGs for 7 d (representative images from n = 8 spheroids). j OVCAR8 spheroid area in collagen-based OHGs after 7 d culture relative to DMSO control ( n = 8 spheroids). k BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion depth into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m OVCAR8 spheroid area quantifications in MMP-cleavable or non-cleavable NB-HA OHG after 7 d culture relative to MMP-cleavable control ( n = 11 spheroids). For the data in ( b , e , h , j and l ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( m ). Error bars in ( b ) represent the s.e.m. Scale bars, 100 μm ( g , k ), 200 μm ( d , f , i ). Source data are provided as a Source Data file.

    Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

    Techniques: Control, Staining

    a F-actin (grey) and DAPI (blue) in OVCAR8 cells embedded in collagen or collagen-based organo-hydrogels (OHG) for 24 h (representative images from n = 3 gels). Microdroplet locations are indicated with an asterisk. b BODIPY (green), F-actin (grey) and nuclei (blue) staining of OVCAR8 cells embedded in collagen-based OHG (left; n = 3 gels) or BODIPY (green), mRFP (red) staining of mRFP-OVCAR8 cells invading into peritoneal adipose tissue (right; n = 3 tissues from 1 donor). Arrowheads indicate oil microdroplet (left) and adipocyte (right) deformations at the contact points with cells. c Quantification of OVCAR8 organotypic invasion depth into peritoneal adipose tissues after 7 d ( n = 3 tissues from 1 donor). Rhombuses indicate the average. d Schematic representation of the Sylgard 184 PDMS-based OGH. e F-actin (grey) and nuclei (red) staining of OVCAR8 spheroids embedded for 7 d in collagen-based OHGs prepared with PDMS microdroplets of varying stiffness (representative images from n = 10 spheroids). f OVCAR8 spheroid area after 7 d in collagen-based OHGs with PDMS microdroplets prepared with varying crosslinker percentage, relative to average area in 2 wt% microbead OHGs ( n = 10 spheroids). g Schematic representation of the norbornene-functionalised hyaluronic acid (NB-HA) OGH with norbornene-functionalised bovine serum albumin (NB-BSA) used for RGD presentation at the microdroplet surface. h F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHG with localised presentation of RGD (left). Relative OVCAR8 spheroid area in NB-HA OHG with localised RGD presentation after 7 d in culture ( n = 11 spheroids; top right). BODIPY (green), nuclei (red), and F-actin (grey) staining of OVCAR8 cells in NB-HA OHG presenting RGD on the microdroplet surface (bottom right). Arrowhead indicates a cell spreading at the microdroplet-HA interface. i Schematic representation of the system to modulate microdroplet interfacial mechanics. j F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHGs (microdroplet-restricted RGD presentation) with varying microdroplet interfacial modulus (representative images from n = 9 spheroids). k OVCAR8 spheroid area in NB-HA OHGs with microdroplet-presenting RGD and varying protein nanosheet interfacial mechanics after 7 d in culture, relative to BSA-only control ( n = 9 spheroids). For the data in ( c and h ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( f and k ). Scale bars, 25 μm ( a , b ), 50 μm ( h , bottom right), 200 μm ( e , h left, j ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

    doi: 10.1038/s41467-025-62296-7

    Figure Lengend Snippet: a F-actin (grey) and DAPI (blue) in OVCAR8 cells embedded in collagen or collagen-based organo-hydrogels (OHG) for 24 h (representative images from n = 3 gels). Microdroplet locations are indicated with an asterisk. b BODIPY (green), F-actin (grey) and nuclei (blue) staining of OVCAR8 cells embedded in collagen-based OHG (left; n = 3 gels) or BODIPY (green), mRFP (red) staining of mRFP-OVCAR8 cells invading into peritoneal adipose tissue (right; n = 3 tissues from 1 donor). Arrowheads indicate oil microdroplet (left) and adipocyte (right) deformations at the contact points with cells. c Quantification of OVCAR8 organotypic invasion depth into peritoneal adipose tissues after 7 d ( n = 3 tissues from 1 donor). Rhombuses indicate the average. d Schematic representation of the Sylgard 184 PDMS-based OGH. e F-actin (grey) and nuclei (red) staining of OVCAR8 spheroids embedded for 7 d in collagen-based OHGs prepared with PDMS microdroplets of varying stiffness (representative images from n = 10 spheroids). f OVCAR8 spheroid area after 7 d in collagen-based OHGs with PDMS microdroplets prepared with varying crosslinker percentage, relative to average area in 2 wt% microbead OHGs ( n = 10 spheroids). g Schematic representation of the norbornene-functionalised hyaluronic acid (NB-HA) OGH with norbornene-functionalised bovine serum albumin (NB-BSA) used for RGD presentation at the microdroplet surface. h F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHG with localised presentation of RGD (left). Relative OVCAR8 spheroid area in NB-HA OHG with localised RGD presentation after 7 d in culture ( n = 11 spheroids; top right). BODIPY (green), nuclei (red), and F-actin (grey) staining of OVCAR8 cells in NB-HA OHG presenting RGD on the microdroplet surface (bottom right). Arrowhead indicates a cell spreading at the microdroplet-HA interface. i Schematic representation of the system to modulate microdroplet interfacial mechanics. j F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHGs (microdroplet-restricted RGD presentation) with varying microdroplet interfacial modulus (representative images from n = 9 spheroids). k OVCAR8 spheroid area in NB-HA OHGs with microdroplet-presenting RGD and varying protein nanosheet interfacial mechanics after 7 d in culture, relative to BSA-only control ( n = 9 spheroids). For the data in ( c and h ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( f and k ). Scale bars, 25 μm ( a , b ), 50 μm ( h , bottom right), 200 μm ( e , h left, j ). Source data are provided as a Source Data file.

    Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

    Techniques: Staining, Control

    a Schematic representation of the organo-hydrogels (OHGs) with varying microdroplet diameter and volume fraction. b Storage modulus of collagen-based OHG with distinct microdroplet size ( n = 4 gels; average ± s.e.m.). The dashed line shows the predicted trend. c Normalised stress-relaxation curves of collagen-based OHG with distinct microdroplet size ( n = 4 gels). d BODIPY (green) and F-actin (red) and nuclei (blue) staining of OVCAR8 cells after 7 d in collagen-based OHG of 70 or 25 μm diameter microdroplets (representative images from n = 13 spheroids). Arrowheads indicate cells at the invasive front. e Quantification of relative OVCAR8 spheroid area in collagen-based OHG of distinct emulsion percentage of volume fraction and microdroplet diameter after 7 d in culture ( n = 13 spheroids). f BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells after 7 d invasion into adipose peritoneal tissue explants (representative images from n = 3 explants per donor). g Correlation between patient average adipocyte diameter and invaded OVCAR8 cells after 7 d, at 42 μm tissue depth, relative to number of cells at the tissue surface ( n = 6 donors; average ± s.e.m.). The dashed line shows the predicted trend. h Quantification of interdroplet distance in OHG ( n = 100 microdroplets). i Quantification of the percentage of invasive front OVCAR8 cells in contact with microdroplets in OHG ( n = 121 cells in 3 gels). j Quantification of percentage of Ki67 + cells, immunofluorescence signal intensity of cytoplasmic pFAK and pMLC relative to collagen average, and nuclear:cytoplasmic ratio of YAP/TAZ in OVCAR8 cells embedded in OHG of 70 or 25 μm diameter microdroplets for 24 h ( n = 55 cells). k F-actin (green) and DAPI (blue) staining of CAOV3 spheroids embedded for 7 d in collagen-based 25 μm microdroplet OHG in the presence or absence of TGFβ. Arrowheads indicate invading cells (representative images from n = 6 spheroids). l Quantification of CAOV3 spheroid area relative to non-TGFβ-treated control after 7 d in culture ( n = 6 spheroids). m Schematic illustration of the cell force-dependent invasion of adipose tissue enabled by the anisotropic mechanics of OHGs and the generation of migration tracks at the ECM-adipocyte/microdroplet interface. Arrows indicate the direction of the force. For the data in ( h , i , j and l ), a two-sided unpaired t test was performed. Coefficient of determination and Pearson correlation (two-tailed test) were performed in ( g ) to determine the relationship between tissue adipocyte diameter and OVCAR8 invasion. Error bars in ( b ) represent the s.e.m. Scale bars, 50 μm (bottom panels in d , k ), 100 μm ( f ), 200 μm (top panels in d , k ). Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

    doi: 10.1038/s41467-025-62296-7

    Figure Lengend Snippet: a Schematic representation of the organo-hydrogels (OHGs) with varying microdroplet diameter and volume fraction. b Storage modulus of collagen-based OHG with distinct microdroplet size ( n = 4 gels; average ± s.e.m.). The dashed line shows the predicted trend. c Normalised stress-relaxation curves of collagen-based OHG with distinct microdroplet size ( n = 4 gels). d BODIPY (green) and F-actin (red) and nuclei (blue) staining of OVCAR8 cells after 7 d in collagen-based OHG of 70 or 25 μm diameter microdroplets (representative images from n = 13 spheroids). Arrowheads indicate cells at the invasive front. e Quantification of relative OVCAR8 spheroid area in collagen-based OHG of distinct emulsion percentage of volume fraction and microdroplet diameter after 7 d in culture ( n = 13 spheroids). f BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells after 7 d invasion into adipose peritoneal tissue explants (representative images from n = 3 explants per donor). g Correlation between patient average adipocyte diameter and invaded OVCAR8 cells after 7 d, at 42 μm tissue depth, relative to number of cells at the tissue surface ( n = 6 donors; average ± s.e.m.). The dashed line shows the predicted trend. h Quantification of interdroplet distance in OHG ( n = 100 microdroplets). i Quantification of the percentage of invasive front OVCAR8 cells in contact with microdroplets in OHG ( n = 121 cells in 3 gels). j Quantification of percentage of Ki67 + cells, immunofluorescence signal intensity of cytoplasmic pFAK and pMLC relative to collagen average, and nuclear:cytoplasmic ratio of YAP/TAZ in OVCAR8 cells embedded in OHG of 70 or 25 μm diameter microdroplets for 24 h ( n = 55 cells). k F-actin (green) and DAPI (blue) staining of CAOV3 spheroids embedded for 7 d in collagen-based 25 μm microdroplet OHG in the presence or absence of TGFβ. Arrowheads indicate invading cells (representative images from n = 6 spheroids). l Quantification of CAOV3 spheroid area relative to non-TGFβ-treated control after 7 d in culture ( n = 6 spheroids). m Schematic illustration of the cell force-dependent invasion of adipose tissue enabled by the anisotropic mechanics of OHGs and the generation of migration tracks at the ECM-adipocyte/microdroplet interface. Arrows indicate the direction of the force. For the data in ( h , i , j and l ), a two-sided unpaired t test was performed. Coefficient of determination and Pearson correlation (two-tailed test) were performed in ( g ) to determine the relationship between tissue adipocyte diameter and OVCAR8 invasion. Error bars in ( b ) represent the s.e.m. Scale bars, 50 μm (bottom panels in d , k ), 100 μm ( f ), 200 μm (top panels in d , k ). Source data are provided as a Source Data file.

    Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

    Techniques: Staining, Emulsion, Immunofluorescence, Control, Migration, Two Tailed Test

    CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), OVCAR8 ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.

    Journal: The Journal of Biological Chemistry

    Article Title: Creatine kinase B promotes non–small cell lung cancer survival and metastasis

    doi: 10.1016/j.jbc.2025.110805

    Figure Lengend Snippet: CKB is the predominant active isoenzyme in bone and ovarian cancer. A , cell survival analysis, as determined by SRB staining, of 143B ( n = 3–6 per dose), OVCAR8 ( n = 4–5 per dose), and MDA-MB-231 ( n = 3–6 per dose) cells treated for 3 days with cyclocreatine. B – D , colony formation assays of 143B ( n = 4 per group), OVCAR8 ( n = 3 per group), and MDA-MB-231 ( n = 3 per group) cells. Crystal violet staining was analyzed by ImageJ software. E , relative CKB and CKMT1 mRNA expression in 143B, OVCAR8, and MDA-MB-231 cells ( n = 3 per group). F , relative CKB mRNA expression in 143B cells stably expressing empty vector (EV), sh CKB #2, or sh CKB #3 ( n = 3 per group). G , relative CKB mRNA expression in OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). H , Western blot of 143B and OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3. I , CK activity in protein lysates from 143B cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). J , CK activity in protein lysates from OVCAR8 cells stably expressing EV, sh CKB #2, or sh CKB #3 ( n = 3 per group). K , cell survival analysis, as determined by SRB staining, of 143B cells treated for 3 days with cyclocreatine ( n = 4 per group). L , cell survival analysis, as determined by SRB staining, of OVCAR8 cells treated for 3 days with cyclocreatine ( n = 4 per group, except for n = 3 for sh CKB #2 and sh CKB #3 at 9 mM and 25 mM). Data are presented as mean ± SEM. n numbers are of biologically independent experiments. B – G , I and J , one-way ANOVA (Dunnett’s multiple comparison test). CK, creatine kinase; CKB , brain-type CK; SRB, Sulforhodamine B.

    Article Snippet: Cell line sources: H1299: American Type Culture Collection (ATCC; CRL-5803); A549: ATCC (CCL-185), OVCAR8: Dr Sidong Huang (McGill University), 143B: ATCC (CRL-8303), and MDA-MB-231: ATCC (CRM-HTB-26).

    Techniques: Staining, Software, Expressing, Stable Transfection, Plasmid Preparation, Western Blot, Activity Assay, Comparison

    ( A ) Gene and pathways enrichment analysis of 1Ai-3638 and 1Ai-8214 downregulated genes by the Gene Analytics tool. ( B ) OVCAR8 ovarian carcinoma cell line was incubated with increasing concentrations of 1Ais for 72 h, and then cell viability was assessed. Viability (100%) is normalized to 0 concentration (DMSO), n = 3 independent biological replicates, and error bars represent SD. ( C ) 1Ais toxicity assessment. Mice weight was measured at different time points after 1Ai-3638 and 1Ai-8214 subcutaneous injection for 3 weeks (assessed every 3–4 days, 3 mice per concentration, 21 mice in total) ( D ) OVCAR8 cell-derived xenografts weight after 1Ais treatment (10 mice per group). OVCAR8 cells were injected into the ovaries of immunocompromised female mice. Out of ten mice in each group (1Ai-3638,1Ai-8214 and DMSO) nine were included as they presented an ovarian tumor. 1Ais were injected into the mice’s peritoneal cavity five times a week for 4 weeks. Finally, mice were sacrificed, and tumor weight was measured. Results are presented as boxplots in which the center = median, box ends = interquartile range (IQR), bottom whisker = minimum and upper whisker= maximum, n = 9. Weight monitoring of mice during the experiment is presented in Appendix Fig. S . * P < 0.05, ** P < 0.01, Student t test. .

    Journal: The EMBO Journal

    Article Title: Inhibitors of eIF1A-ribosome interaction unveil uORF-dependent regulation of translation initiation and antitumor and antiviral effects

    doi: 10.1038/s44318-025-00449-6

    Figure Lengend Snippet: ( A ) Gene and pathways enrichment analysis of 1Ai-3638 and 1Ai-8214 downregulated genes by the Gene Analytics tool. ( B ) OVCAR8 ovarian carcinoma cell line was incubated with increasing concentrations of 1Ais for 72 h, and then cell viability was assessed. Viability (100%) is normalized to 0 concentration (DMSO), n = 3 independent biological replicates, and error bars represent SD. ( C ) 1Ais toxicity assessment. Mice weight was measured at different time points after 1Ai-3638 and 1Ai-8214 subcutaneous injection for 3 weeks (assessed every 3–4 days, 3 mice per concentration, 21 mice in total) ( D ) OVCAR8 cell-derived xenografts weight after 1Ais treatment (10 mice per group). OVCAR8 cells were injected into the ovaries of immunocompromised female mice. Out of ten mice in each group (1Ai-3638,1Ai-8214 and DMSO) nine were included as they presented an ovarian tumor. 1Ais were injected into the mice’s peritoneal cavity five times a week for 4 weeks. Finally, mice were sacrificed, and tumor weight was measured. Results are presented as boxplots in which the center = median, box ends = interquartile range (IQR), bottom whisker = minimum and upper whisker= maximum, n = 9. Weight monitoring of mice during the experiment is presented in Appendix Fig. S . * P < 0.05, ** P < 0.01, Student t test. .

    Article Snippet: OVCAR8 , Creative Biolabs , IOC-ZP305.

    Techniques: Incubation, Concentration Assay, Injection, Derivative Assay, Whisker Assay