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 ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn.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

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

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

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

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

Journal: Cell Reports Medicine

Article Title: Early-stage multi-cancer detection through a plasma extracellular vesicle protein signature

doi: 10.1016/j.xcrm.2026.102694

Figure Lengend Snippet: Transformation-induced changes to the protein composition of cell-derived sEVs (A) The morphology of isolated sEVs was assessed using transmission electron microscopy. Images of normal and transformed HBEC-derived sEVs (scale bars, 200 nm). (B) Nanoparticle analysis using tunable resistive pulse sensing of sEVs isolated from HBECs demonstrates that the majority of sEVs have a size range between 30 and 150 nm, and that transformation does not result in an increase in sEV secretion. (C) Western blot of sEVs from HBECs demonstrating the presence of sEV proteins HSP70 and CD63 and the absence of the cell marker calnexin. (D) Label-free mass spectrometry identified 148 proteins with greater abundance in sEVs derived from transformed HBECs (FDR <0.02), of which 15 were annotated as extracellular proteins. (E) Mass spectrometry results were confirmed using ELISA for THBS1, NID1, PTX3, and VCAN in sEVs derived from normal and transformed HBECs. (F) sEVs derived from 22 cancer cell lines including NSCLC (SKMES1, H1650, HCC4006, and H2170), glioblastoma ([GBM], D54, D270, U87, and U118), colorectal cancer ([CRC], HT29 and SW620), breast cancer ([BCa], BT549, MDA231, and MDA436), prostate cancer ([PCa], PC3 and LNCaP), melanoma ([MEL], A375, MAMEL65, and SKMEL28), esophageal cancer ([ECa], OE19), and ovarian cancer ([OVA], A2780, CAOV3, IGROV1, and OVCAR8) showed a clear increase in expression of THBS1, NID1, PTX3, and VCAN in relation to the average levels of sEVs from normal cells ([HBEC] 30KT, HOSE 6.3, and HOSE 17.1). Samples in mass spectrometry and ELISA were measured in triplicate. See also and .

Article Snippet: OVCAR8 , Creative Biolabs , Cat# IOC-ZP305; RRID: CVCL_1629.

Techniques: Transformation Assay, Derivative Assay, Isolation, Transmission Assay, Electron Microscopy, Tunable Resistive Pulse Sensing, Western Blot, Marker, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: Overexpression of lncRNA LINC00857 in ovarian cancer cells and tissues. ( A ) Relative expression levels of LINC00857 in OC tissues compared to adjacent normal tissues using qRT-PCR ( n = 50). ( B ) Relative expression levels of LINC00857 in OC cell line OVCAR8 compared to normal ovarian epithelial cell line IOSE80 using qRT-PCR ( n = 25). Data were presented as mean ± SD; ** p < 0.01; statistical significance was determined using unpaired Student’s t-test. OC, ovarian cancer.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Over Expression, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: LINC00857 enhances the invasion, migration, proliferation, and stemness of ovarian cancer cell OVCAR8. ( A ) qRT-PCR analysis of LINC00857 expression levels in OVCAR8 cells transfected with si-NC, si-LINC00857, the vector group, or a LINC00857 overexpression plasmid. ( B ) Cell viability was assessed using CCK-8 assay. ( C ) Cell invasion was evaluated using Transwell assay (scale bar = 50 μm). ( D ) Cell migration was measured using scratch assay (scale bar = 100 μm). ( E ) Representative images of sphere-formation assay showing the sphere-forming capacity of OVCAR8 cells in each group (scale bar = 50 μm). ( F–G ) The protein levels of SOX2, NANOG, and Oct4 in OVCAR8 cells. Data were presented as mean ± SD ( n = 3 ~ 5); ** p < 0.01 vs. si-NC; ## p < 0.01 vs. Vector; statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. CCK-8, Cell Counting Kit-8; SOX2, SRY-box transcription factor 2; NANOG, Nanog homeobox; Oct4, octamer-binding transcription factor 4.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Migration, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Over Expression, CCK-8 Assay, Transwell Assay, Wound Healing Assay, Tube Formation Assay, Cell Counting, Binding Assay

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: LINC00857 participates in the activation of YAP1. ( A ) Correlation analysis between LINC00857 and YAP1 expression based on TCGA database ( n = 353). ( B ) Immunohistochemistry analysis of YAP1 levels in OC tissues compared to adjacent tissues (scale bar = 20 μm); ** p < 0.01. ( C ) Western blot analysis of YAP1 protein expression levels in IOSE80 and OVCAR8 cells; ** p < 0.01. ( D ) Western blot analysis of YAP1, p-YAP1, LATS1, p-LATS1, and TEAD4 protein levels in OVCAR8 cells across groups. Data were presented as mean ± SD ( n = 3); ** p < 0.01 vs. si-NC; ## p < 0.01 vs. Vector; statistical analysis was performed using unpaired two-tailed Student’s t-test (for two-group comparisons) or one-way ANOVA followed by Tukey’s post hoc test (for multiple comparisons). YAP1, yes-associated protein 1; TCGA, The Cancer Genome Atlas; OC, ovarian cancer; p-YAP1, phosphorylated-YAP1; LATS1, large tumor suppressor 1; p-LATS1, phosphorylated-LATS1; TEAD4, TEA domain transcription factor 4.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Activation Assay, Expressing, Immunohistochemistry, Western Blot, Plasmid Preparation, Two Tailed Test

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: METTL3 highly expresses in ovarian cancer cells and promotes the LINC00857 expression by m6A modification. ( A ) Relative expression levels of METTL3 in OC tissues compared to adjacent normal tissues using qRT-PCR ( n = 50). ( B ) The correlation between LINC00857 and METTL3 expression analyzed by Pearson ( n = 108). ( C ) Relative expression levels of METTL3 in OC cell line OVCAR8 compared to normal ovarian epithelial cell line IOSE80 using qRT-PCR ( n = 25). ( D ) The m6A enrichment levels of LINC00857 in IOSE80 and OVCAR8 cells were measured using the MeRIP-qPCR kit ( n = 25). ( E ) Relative METTL3 mRNA expression levels in OVCAR8 cells transfected with si-METTL3 or si-NC ( n = 5). ( F ) Western blot analysis showing METTL3 protein expression in OVCAR8 cells transfected with si-NC or si-METTL3 ( n = 3). ( G ) Relative mRNA expression of LINC00857 in OVCAR8 cells after METTL3 knockdown (si-METTL3) compared to the si-NC group ( n = 5). ( H ) m6A enrichment levels of LINC00857 in OVCAR8 cells transfected with si-METTL3 or si-NC using MeRIP-qPCR ( n = 5). ( I ) RNA pull-down assay showing the binding interaction between LINC00857 and METTL3 ( n = 5). ( J ) RNA stability assay indicating the relative expression of LINC00857 over time in OVCAR8 cells transfected with si-METTL3 or si-NC ( n = 5). Data were presented as mean ± SD; ** p < 0.01; statistical significance was determined using unpaired Student’s t-test. METTL3, methyltransferase-like 3; OC, ovarian cancer; m6A, N6-methyladenosine.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Expressing, Modification, Quantitative RT-PCR, Transfection, Western Blot, Knockdown, Pull Down Assay, Binding Assay, Stability Assay

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: LINC00857 reverses the migration, invasion, proliferation, and stemness of OVCAR8 cells inhibited by METTL3 down-regulation. ( A ) Cell viability of OVCAR8 cells was assessed using CCK-8 assay. ( B ) Cell invasion was evaluated using Transwell assay (scale bar = 50 μm). ( C ) Cell migration was measured using scratch assay (scale bar = 100 μm). ( D ) Representative images of sphere-formation assays showing the sphere-forming capability of OVCAR8 cells in each group (scale bar = 50 μm). ( E–F ) Western blot analysis showing the protein levels of SOX2, Oct4, and NANOG in OVCAR8 cells. Data were presented as mean ± SD ( n = 3 ~ 5); ** p < 0.01 vs. NC; ## p < 0.01 vs. si-METTL3; statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. METTL3, methyltransferase-like 3; CCK-8, Cell Counting Kit-8; SOX2, SRY-box transcription factor 2; NANOG, Nanog homeobox; Oct4, octamer-binding transcription factor 4.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Migration, CCK-8 Assay, Transwell Assay, Wound Healing Assay, Western Blot, Cell Counting, Binding Assay

Journal: Scientific Reports

Article Title: METTL3-mediated m6A modification of LINC00857 enhances stemness and metastasis of ovarian cancer cells by activating the YAP-TEAD pathway

doi: 10.1038/s41598-025-24958-w

Figure Lengend Snippet: LINC00857 reverses the effect of down-regulation of METTL3 on YAP1. ( A ) Representative protein bands of YAP1, p-YAP1, LATS1 and TEAD4 in OVCAR8 cells across groups. ( B ) Quantification of protein expression levels normalized to GAPDH. Data were presented as mean ± SD ( n = 3); ** p < 0.01 vs. NC; ## p < 0.01 vs. si-METTL3; statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. METTL3, methyltransferase-like 3; YAP1, yes-associated protein 1; p-YAP1, phosphorylated-YAP1; LATS1, large tumor suppressor 1; TEAD4, TEA domain transcription factor 4.

Article Snippet: OC cells (OVCAR8) were provided by Procell Life Science&Technology Co., Ltd (CL-0178, China).

Techniques: Expressing

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