Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: Specific IFN-γ and TNF-α release of T lymphocytes transduced with TIM-3-silenced HER2-specific chimeric antigen receptor (CAR) or HER2-specific CAR. (A, B) TIM-3-silenced CAR-T cells and control T cells were co-incubated with Galectin-9 + or Galectin-9 – SKOV3 tumor cells (E:T ratio 5:1 or 10:1). At 20 h after coculture, a specific enzyme-linked immunosorbent assay was used to analyze the supernatant for IFN-γ cytokine-release. Results were presented as mean ± standard deviation. (C, D) The detection of TNF-α in the same culture supernatant. Results were presented as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Transduction, Control, Incubation, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Genes & Diseases

Article Title: Blockade of co-inhibitory receptor immune checkpoint protein TIM3/CD366 augments the anti-cancer activity of CAR-T therapy in solid tumors: An ovarian cancer example

doi: 10.1016/j.gendis.2025.101978

Figure Lengend Snippet: TIM-3 silencing augmented the anti-tumor activity of chimeric antigen receptor-T (CAR-T) cells in vivo . 2 × 10 6 SKOV3 tumor cells expressing luciferase were intraperitoneally inoculated in a xenograft mouse model, and 7 days after inoculation, the 2 × 10 6 HER2-specific CAR-T kdTim-3 cells or CAR-T cells, or untreated T cells were intraperitoneally administered. (A, B) Tumor growth was monitored using an in vivo imaging system. (C) Survival curve of 80-day post-treatment. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: Human cervical cancer cell line HeLa, lentivirus packaging cell line HEK 293TD, and human ovarian cancer cell line SKOV3 were purchased from American Type Culture Collection (Manassas, Virginia, USA) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, Grand Island, New York) supplemented with 10% heat-inactivated fetal bovine serum.

Techniques: Activity Assay, In Vivo, Expressing, Luciferase, In Vivo Imaging

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: Native NPM1 and YAP1 proteins form complexes in prostate cancer cell models (A) The Venn diagram illustrates proteins associated with YAP1 in LNCaP cells treated with dihydrotestosterone (DHT, 10 nM) or enzalutamide (ENZ, 20 μM) under dextran-coated charcoal-stripped (DCC) serum conditions. Unique and shared proteins for each treatment are indicated. (B) A protein interaction network generated using the GeneMania web portal positions NPM1 within the YAP1 network. (C) Western blot analysis of NPM1 protein levels in AR-positive (LNCaP, C4-2, C4-2B, and 22Rv1) and AR-negative (PC3 and ARCaP) cell lines . β-actin served as a loading control. (D) Quantitative PCR analysis showing correlation between NPM1 transcript and protein levels across the same cell lines. (E) Co-immunofluorescence staining of NPM1 (green) and YAP1 (red) in LNCaP and PC3 cells; nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. (F) Western blot showing NPM1 in YAP1 immune complexes in nuclear extracts from LNCaP, C4-2, C4-2B, and PC3 cells. IgG served as a negative control . (G) Reciprocal co-IP using an NPM1 antibody demonstrates YAP1 presence in NPM1 immune complexes exclusively in PC3 cells. β-actin was used as a loading control. (H) Representative confocal images showing PLA foci (red) indicating YAP1-NPM1 interactions in LNCaP, C4-2, and PC3 cells; nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. (I) Enlarged images of boxed regions from panel H highlight nuclear localization of PLA foci. (J) Quantification of PLA foci per cell across the indicated cell lines. Statistical significance was determined using a two-tailed t test (∗ p < 0.01). Data are represented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Generated, Western Blot, Control, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Negative Control, Co-Immunoprecipitation Assay, Two Tailed Test

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: NPM1 influences YAP1 activity and the regulation of its target genes in cellular contexts (A–C) Quantitative PCR analysis of NPM1, YAP1, and MYC mRNA levels in LNCaP and PC3 cells after transient transfection with scrambled or NPM1-specific siRNA. (D and E) Western blot analysis showing YAP1 and MYC protein levels in LNCaP and PC3 cells with or without NPM1 knockdown. (F–G) Quantitative PCR analysis of YAP1 target gene expression in LNCaP and PC3 cells transfected with scramble control or NPM1-specific siRNA (∗ p < 0.001; ∗∗ p < 0.01). Data are represented as mean ± SEM. (I–K) Quantitative PCR analysis of YAP1 target genes (CCN1, CCN2, and ANKRD1) in LNCaP cells after transfection with scramble control or NPM1-specific siRNA, followed by overnight treatment with EtOH (vehicle) or DHT under 5% CSS-fed conditions . Statistical significance was assessed using a two-tailed t test (∗ p < 0.05; ∗∗ p < 0.01). Data are presented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Knockdown, Targeted Gene Expression, Control, Two Tailed Test

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: Synergistic interactions between NPM1 and YAP1 regulate cell growth (A) Western blot analysis of NPM1 protein levels in LNCaP cells with or without NPM1 silencing by siRNA; β-actin served as a loading control. (B) Colony-forming ability of LNCaP cells with or without NPM1 knockdown, visualized by crystal violet staining. The accompanying graph presents quantification of colony numbers (∗ p < 0.01, two-tailed t test). (C and D) Dose-response curves for LNCaP, C4-2, C4-2B, and PC3 cells treated with NPM1 inhibitors NSC348884 or nucleozin. Graphs show log values of drug concentration versus percent cell growth; IC50 values were calculated for each cell line. (E–G) Cell-cycle distribution in LNCaP, C4-2, and PC3 cells treated with DMSO (mock), NSC348884, or nucleozin, as evaluated by flow cytometry. Graphs display the percentages of cells in the G1, S, and G2/M phases. (H) Confocal images showing YAP1–NPM1 interactions in LNCaP cells treated with DMSO, NSC348884, or nucleozin under serum-fed conditions. PLA was used to detect YAP1-NPM1 interactions (red foci); nuclei were counterstained with DAPI (blue). Scale bar: 10 μm. The accompanying graph quantifies PLA foci. (I) Cell growth with or without YAP1 induction under NPM1 knockdown conditions, assessed by CCK-8 assay at 72 h post-transfection (∗ p < 0.001; ∗∗ p < 0.01, two-tailed t test). Data are presented as mean ± SEM. (I) Cell growth with or without YAP1 induction under NPM1 knockdown conditions, assessed by CCK-8 assay at 72 h post-transfection (∗ p < 0.001; ∗∗ p < 0.01, two-tailed t test). Data are represented as mean ± SEM.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Control, Knockdown, Staining, Two Tailed Test, Concentration Assay, Flow Cytometry, CCK-8 Assay, Transfection

Journal: iScience

Article Title: The YAP1-NPM1 nuclear complex regulates MYC and reveals a targetable oncogenic node

doi: 10.1016/j.isci.2026.115588

Figure Lengend Snippet: NPM1 regulates cell migration and shows increased interaction with YAP1 in high-grade prostate cancer tissues (A) Wound-healing assay images demonstrate a time-dependent delay in wound closure in LNCaP cells following NPM1 knockdown compared to scramble (Scram) siRNA control ( p < 1.14E−13). Scale bar: 10 μm. (B and C) Migration assays in LNCaP and PC3 cells show reduced motility upon NPM1 depletion in collagen-coated Boyden chambers, with or without EGF stimulation (∗ p < 0.01). Scale bar: 10 μm. Data are from two independent experiments performed in duplicate. (D) Representative PLA micrographs reveal increased YAP1-NPM1 interaction in high-grade tumors (GS > 7) versus low-grade (GS < 7) prostate cancer tissues ( n = 8). Scale bars: 10 and 5 μm, respectively. (E) Quantification of PLA foci in red (corresponding to D) ( n = 8). Data are expressed as mean ± SEM. (F) Schematic summary of findings created using BioRender.

Article Snippet: LNCaP (Cat# CRL-1740), C4-2 (Cat# CRL-3314), C4-2B (Cat# CRL-3315), 22Rv1 (Cat# CRL-2505), and PC3 (Cat# CRL-1435) cell lines were obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, Wound Healing Assay, Knockdown, Control

Journal: Research

Article Title: The Selective DHCR24 Blocker SH42 Inhibits ACE2 Binding and Cellular Entry of SARS-CoV-2 Spike Proteins More Efficiently Than Atorvastatin

doi: 10.34133/research.1280

Figure Lengend Snippet: SH42 reduces cholesterol abundance in the plasma membrane in general and lipid rafts in particular, and decreases lipid raft area more efficiently than atorvastatin (ATO). Control HEK293T (A) and Calu-3 (B) cells and those treated for 96 h with 10 nM or 1 μM ATO or SH42 were labeled with cholesterol-binding mCherry-conjugated D4H*, the D434S mutant of domain 4 (D4) of C. perfringens theta-toxin. Fluorescence intensities correlating with plasma membrane cholesterol levels of at least 10,000 individual cells per sample were subsequently measured using a flow cytometer. The average intensity values obtained in n = 9 independent biological replicates, and their average values (± SEM) are plotted in the figure. (C) To examine changes in the cholesterol content of raft and non-raft microdomains of the plasma membrane, control HEK/ACE2 + TMPRSS2 cells and those treated as above were labeled with Alexa Fluor 647-conjugated cholera toxin subunit B (CTX-AF647), a lipid raft marker, and F66. F66 is a fluorescent indicator with spectral properties depending on the cholesterol-dependent local molecular order (dipole potential) of the membrane; therefore, this dye, combined with CTX-AF647, can provide information about the extent of cholesterol reduction separately in raft and non-raft membrane regions. Representative confocal microscopic images taken from the flat, bottom membrane region adjacent to the coverglass show F66 intensities detected in 2 wavelength ranges of emission (“F66 N*” and “F66 T*”), their ratio (“F66 emission ratio” calculated as T*/N* pixel by pixel), and CTX-AF647 intensities. Cell “membrane masks” selected manually in CTX images were segmented using the maxentropy algorithm to CTX-high “rafts” and CTX-low “non-rafts” corresponding to high- and low-intensity regions, respectively, as shown by the representative images. Violin plots were generated from median F66 emission ratio values determined separately for the CTX-high “raft” (D) and CTX-low “non-raft” (E) masks of n = 54 to 73 individual cells, which also display median values with quartiles. (F) Pixelwise distributions of the F66 emission ratio in CTX-high “rafts” and CTX-low “non-rafts” of control cells are displayed. For the quantification of the relative area of lipid rafts, as an alternative definition for raft regions, a threshold value of the F66 emission ratio was determined (green dashed line) and membrane pixels were considered as “F66 raft” and “F66 non-raft” regions when being above and below the threshold, respectively. (G) Violin plots were generated from the relative fraction of F66 raft pixels (“F66 raft area”) of individual cells, which also display median values with quartiles. (H) Representative images show changes in the lateral distribution of the F66 emission ratio on a color-scale image and reduction in the relative F66 raft area induced by 1 μM SH42. Throughout the figure, asterisks indicate significant differences compared to control samples (* P < 0.05, **** P < 0.0001), while hashes show that between samples treated with ATO and SH42 at identical concentrations ( # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001), which were determined by Tukey’s HSD test carried out after significant differences were obtained for between-group effects in ANOVA.

Article Snippet: The human embryonic kidney HEK293T cell line that stably expresses ACE2 and transmembrane serine protease 2 (TMPRSS2) genes (HEK/ACE2 + TMPRSS2) was obtained from GeneCopoeia (Rockville, MD; SL222), while the Calu-3 lung adenocarcinoma cell line with an endogenous expression of ACE2 and TMPRSS2, and the original HEK293T cell line lacking considerable ACE2 and TMPRSS2 were purchased from the American Type Culture Collection (Manassas, VA; HTB-55 and CRL-3216, respectively).

Techniques: Clinical Proteomics, Membrane, Control, Labeling, Binding Assay, Mutagenesis, Fluorescence, Flow Cytometry, Marker, Generated

Journal: Research

Article Title: The Selective DHCR24 Blocker SH42 Inhibits ACE2 Binding and Cellular Entry of SARS-CoV-2 Spike Proteins More Efficiently Than Atorvastatin

doi: 10.34133/research.1280

Figure Lengend Snippet: SH42 decreases ACE2 binding of SARS-CoV-2 spike receptor-binding domains (RBDs) more efficiently than ATO. (A) ACE2-expressing HEK/ACE2 + TMPRSS2 and Calu-3 control cells and those treated for 96 h with 10 nM or 1 μM ATO or SH42 were incubated with the GFP-conjugated RBDs of the Wuhan-Hu-1 strain (WT), Delta, and Omicron BA.1 variants for 4 min. RBDs were applied at 0.2 and 1.0 μg/ml for HEK/ACE2 + TMPRSS2 and Calu-3 cells, respectively. Fluorescence intensities of at least 10,000 individual cells per sample were subsequently measured using a flow cytometer. (B) Representative RBD-GFP versus forward-scattered light intensity (FSC) density plots demonstrate decreases in the bound WT RBD-GFP in response to 1 μM SH42 in HEK/ACE2 + TMPRSS2 cells. Dashed lines represent average values of the fluorescence intensity obtained in the displayed representative samples. The average intensities obtained in n = 9 independent biological replicates and normalized to the mean value determined in untreated control samples, and their average values (± SEM) are plotted for WT, Delta, and Omicron BA.1 variants in HEK/ACE2 + TMPRSS2 (C) and Calu-3 (D) cells. Throughout the figure, asterisks indicate significant differences compared to control samples (* P < 0.05, ** P < 0.01, **** P < 0.0001), while hashes show those between samples treated with ATO and SH42 at identical concentrations ( ### P < 0.001, #### P < 0.0001), which were determined by Tukey’s HSD test carried out after significant differences were obtained for between-group effects in ANOVA.

Article Snippet: The human embryonic kidney HEK293T cell line that stably expresses ACE2 and transmembrane serine protease 2 (TMPRSS2) genes (HEK/ACE2 + TMPRSS2) was obtained from GeneCopoeia (Rockville, MD; SL222), while the Calu-3 lung adenocarcinoma cell line with an endogenous expression of ACE2 and TMPRSS2, and the original HEK293T cell line lacking considerable ACE2 and TMPRSS2 were purchased from the American Type Culture Collection (Manassas, VA; HTB-55 and CRL-3216, respectively).

Techniques: Binding Assay, Expressing, Control, Incubation, Fluorescence, Flow Cytometry

Journal: Research

Article Title: The Selective DHCR24 Blocker SH42 Inhibits ACE2 Binding and Cellular Entry of SARS-CoV-2 Spike Proteins More Efficiently Than Atorvastatin

doi: 10.34133/research.1280

Figure Lengend Snippet: SH42-induced reduction in ACE2 binding of WT SARS-CoV-2 spike RBDs negatively correlates with the applied RBD concentration. ACE2-expressing HEK/ACE2 + TMPRSS2 (A) and Calu-3 (B) control cells and those treated for 96 h with 10 nM or 1 μM ATO or SH42 were incubated with different concentrations of the GFP-conjugated RBDs of the Wuhan-Hu-1 strain (WT) for 4 min. Fluorescence intensities of at least 10,000 individual cells per sample were subsequently measured using a flow cytometer. The extents of inhibition (calculated as 1 − average of treated/average of control) were determined in n = 9 independent biological replicates, and their average values (± SEM) are plotted as a function of the applied RBD concentration ranging between 0.1 and 5 μg/ml for HEK/ACE2 + TMPRSS2 and between 1 and 10 μg/ml for Calu-3 cells. Asterisks indicate significant differences between samples treated with the lowest versus highest RBD concentrations for each treatment (** P < 0.01, **** P < 0.0001), which were determined by Tukey’s HSD test carried out after significant differences were obtained for between-group effects in ANOVA.

Article Snippet: The human embryonic kidney HEK293T cell line that stably expresses ACE2 and transmembrane serine protease 2 (TMPRSS2) genes (HEK/ACE2 + TMPRSS2) was obtained from GeneCopoeia (Rockville, MD; SL222), while the Calu-3 lung adenocarcinoma cell line with an endogenous expression of ACE2 and TMPRSS2, and the original HEK293T cell line lacking considerable ACE2 and TMPRSS2 were purchased from the American Type Culture Collection (Manassas, VA; HTB-55 and CRL-3216, respectively).

Techniques: Binding Assay, Concentration Assay, Expressing, Control, Incubation, Fluorescence, Flow Cytometry, Inhibition

Journal: Research

Article Title: The Selective DHCR24 Blocker SH42 Inhibits ACE2 Binding and Cellular Entry of SARS-CoV-2 Spike Proteins More Efficiently Than Atorvastatin

doi: 10.34133/research.1280

Figure Lengend Snippet: SH42 inhibits the cellular entry of SARS-CoV-2 spike trimers more efficiently than ATO. Control HEK/ACE2 + TMPRSS2 and Calu-3 cells and those treated for 96 h with 10 nM or 1 μM ATO or SH42 were incubated for 4 h in the presence of WT, Delta, or Omicron BA.1 SARS-CoV-2 spike trimers conjugated with Alexa Fluor 488 (AF488-trimers) and labeled with F66. (A) Representative orthogonal views of confocal Z-stack images of F66 for the visualization of the plasma membrane and AF488-trimers to estimate entry demonstrate notable trimer accumulation in the intracellular space of untreated control HEK/ACE2 + TMPRSS2 cells. During image analysis, pixels corresponding to plasma membrane and intracellular pixels were segmented based on F66 Z-stack images. Markers were manually placed inside cells (green circles in the grayscale orthogonal view), and a MATLAB implementation of the 3D watershed algorithm identified the intracellular space of cells and their membrane (colored regions and red lines in the orthogonal view in the middle, respectively, and their overlay image displayed on the right). (B) Representative 3D reconstruction images displaying AF488 fluorescence intensities on a green-red color scale above a threshold intensity overlaid on intracellular pixels of individual cells (in transparent blue) demonstrate decreases in the amount of intracellular WT trimers in response to 1 μM SH42. Subsequently, the average fluorescence intensity values emitted by AF488-trimers were calculated exclusively from data of intracellular pixels for individual cells. The average intensities obtained in n = 400 to 600 HEK/ACE2 + TMPRSS2 (C) and Calu-3 (D) cells and normalized to the median value determined in untreated control samples are plotted along with median values with quartiles for WT, Delta, and Omicron BA.1 trimer variants. Asterisks indicate significant differences compared to control samples (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001), while hashes show that between samples treated with ATO and SH42 at identical concentrations ( # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001), which were determined by Tukey’s HSD test carried out after significant differences were obtained for between-group effects in ANOVA. Lognormal functions fitted to normalized mean intracellular AF488-trimer fluorescence intensity histograms of individual HEK/ACE2 + TMPRSS2 (E) and Calu-3 (F) cells also demonstrate the effects of ATO and SH42 on the internalization of WT, Delta, and Omicron BA.1 trimer variants.

Article Snippet: The human embryonic kidney HEK293T cell line that stably expresses ACE2 and transmembrane serine protease 2 (TMPRSS2) genes (HEK/ACE2 + TMPRSS2) was obtained from GeneCopoeia (Rockville, MD; SL222), while the Calu-3 lung adenocarcinoma cell line with an endogenous expression of ACE2 and TMPRSS2, and the original HEK293T cell line lacking considerable ACE2 and TMPRSS2 were purchased from the American Type Culture Collection (Manassas, VA; HTB-55 and CRL-3216, respectively).

Techniques: Control, Incubation, Labeling, Clinical Proteomics, Membrane, Fluorescence

Journal: Research

Article Title: The Selective DHCR24 Blocker SH42 Inhibits ACE2 Binding and Cellular Entry of SARS-CoV-2 Spike Proteins More Efficiently Than Atorvastatin

doi: 10.34133/research.1280

Figure Lengend Snippet: SH42 decreases cell surface ACE2 expression and its colocalization with lipid rafts more efficiently than ATO. (A) Control HEK/ACE2 + TMPRSS2 and Calu-3 cells and those treated for 96 h with 10 nM or 1 μM ATO or SH42 were labeled with Alexa Fluor 488-conjugated anti-ACE2 antibodies (AF488-anti-ACE2). Fluorescence intensities of at least 10,000 individual cells per sample were subsequently measured using a flow cytometer. The average intensities obtained in n = 10 independent biological replicates and normalized to the mean value determined in untreated control samples, and their average values (± SEM) are plotted in the panel. (B) Control cells and those treated as above were labeled with AF488-anti-ACE2 and Alexa Fluor 647-conjugated cholera toxin subunit B (AF647-CTX). Representative confocal microscopic images taken from the flat, bottom membrane region adjacent to the coverglass show AF488-anti-ACE2 and AF647-CTX intensities, and their overlay, while the colocalization of the 2 signals and its changes in response to 1 μM SH42 are displayed in representative dot plots obtained from pixelwise fluorescence intensities. (C) Violin plots were generated from Pearson correlation coefficient values between fluorescence intensities of the 2 applied fluorophores determined from pixelwise data of n = 81 to 90 individual cells, which also display median values with quartiles. Throughout the figure, asterisks indicate significant differences compared to control samples (* P < 0.05, ** P < 0.01, **** P < 0.0001), while hashes show that between samples treated with ATO and SH42 at identical concentrations ( ### P < 0.001, #### P < 0.0001), which were determined by Tukey’s HSD test carried out after significant differences were obtained for between-group effects in ANOVA.

Article Snippet: The human embryonic kidney HEK293T cell line that stably expresses ACE2 and transmembrane serine protease 2 (TMPRSS2) genes (HEK/ACE2 + TMPRSS2) was obtained from GeneCopoeia (Rockville, MD; SL222), while the Calu-3 lung adenocarcinoma cell line with an endogenous expression of ACE2 and TMPRSS2, and the original HEK293T cell line lacking considerable ACE2 and TMPRSS2 were purchased from the American Type Culture Collection (Manassas, VA; HTB-55 and CRL-3216, respectively).

Techniques: Expressing, Control, Labeling, Fluorescence, Flow Cytometry, Membrane, Generated

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin dose‐dependently decreases the growth of ovarian cancer cells. The ovarian cancer cell lines, OVCAR‐3 and SKOV‐3, were treated with PCG (from 6.25 to 200 µM) for 24, 48, and 72 h. Cell proliferation was detected by the WST assay. PCG caused a decrease in the growth of (A–C) OVCAR 3 cells and (D–F) SKOV‐3 cells with varying IC 50 s. IC 50 dose concentrations are presented in (G–I) for OVCAR‐3 cells and (J–L) SKOV‐3 cells. Following the statistical analysis, the data in the graphs have been derived from at least three repeated experiments and shown as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: WST Assay, Derivative Assay, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin suppressed the colony formation ability of ovarian cancer cells. The ovarian cancer cell lines, OVCAR‐3 and SKOV‐3, were treated with PCG (from 6.25 to 200 µM) for 48 h and then incubated for 14 days. PCG causes a decrease in the colony formation ability of the (A) OVCAR‐3 and (B) SKOV‐3 cells. The inhibitory effect was more prominent in the OVCAR‐3 as compared to SKOV‐3 cells.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Incubation

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin decreased the migration ability of ovarian cancer cells. The wound healing assay showed a decrease in the ability of ovarian cancer cells to migrate after PCG treatment. PCG caused such an effect in 48 h in (A) OVCAR‐3 cells and for 24 and 48 h in (B) SKOV‐3 cells. The highest cell migration inhibitory effects were obtained in the SKOV‐3 cells as compared to OVCAR‐3 cells. Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Migration, Wound Healing Assay, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin suppressed the transwell migration ability of ovarian cancer cells. To assess the transwell migration ability of PCG on the ovarian cancer cells, the OVCAR‐3 and SKOV‐3 were subjected to 48 h to PCG at various doses (50, 100, and 200 µM). PCG treatment resulted in the decreased transwell migration ability of the (A) OCAR‐3 and (B) SKOV‐3 cells. (C) Western blot assay revealed a decrease in the N‐cadherin, while an increase in E‐cadherin was observed in SKOV‐3 cancer cells. Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Migration, Western Blot, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin disrupted the mitochondrial membrane potential in ovarian cancer cells. Representative images for the analysis of MMP through the JC‐10 assay in (A) OVCAR‐3 cells and (B) SKOV‐3 cells. All cells were seeded at a density of 4 × 10 3 per well in cell culture dishes and treated with (0.1% DMSO) and PCG (50, 100, and 200 µM). Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Membrane, Cell Culture, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin caused an increase in ROS production in ovarian cancer cells. For the quantification of the ROS, the ovarian cancer cells were subjected to PCG for 48 h before employing the MitoSOX assay. There was an increase in the ROS generation at higher doses for (A) OVCAR‐3 and (B) SKOV‐3 cells. Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Mitosox Assay, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin caused apoptosis in ovarian cancer cells. Representative images for the analysis of Annexin V/PI assay in (A) OVCAR‐3 and (B) SKOV‐3 cells. All cells were seeded at a density of 8 × 10 4 per well in cell culture dishes and treated with (0.1% DMSO) and PCG (50, 100, and 200 µM). (C) Representative image of the BAX expression in the SKOV‐3 cells. Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Cell Culture, Expressing, Control

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Punicalagin Inhibits the Growth and Proliferation of Ovarian Epithelial Adenocarcinoma Cells Via Apoptosis and Autophagic Cell Death

doi: 10.1002/jbt.70908

Figure Lengend Snippet: Punicalagin causes autophagy in ovarian cancer cells. Representative images for the analysis of autophagy through Acridine Orange staining in (A) OVCAR‐3 and (B) SKOV‐3 cells. All cells were seeded at a density of 4 × 10 3 per well in cell culture dishes and treated with (0.1% DMSO) and PCG (50, 100, and 200 µM). Punicalagin caused autophagy in the OVCAR‐3 cell line. (C) Representative image of the BAX expression in the OVCAR‐3 cells. Data were obtained after statistical analysis using at least three repeated experiments, presented in graphs as the mean ± SD. * p ≤ 0.05 compared to control.

Article Snippet: Ovarian cancer cell lines SKOV‐3 and OVCAR‐3 were obtained from ATCC (Manassas, USA).

Techniques: Staining, Cell Culture, Expressing, Control