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: Translational Oncology

Article Title: Integrating spatial and single-cell transcriptomics via machine learning to characterize efferocytosis in hepatocellular carcinoma prognosis and immunotherapy

doi: 10.1016/j.tranon.2026.102801

Figure Lengend Snippet: Identification and multi-level validation of TPI1 as a pivotal prognostic driver. (A) Lollipop chart showing the selection frequency of feature genes across 101 machine learning models, identifying TPI1 as a high-frequency core gene. (B) Univariate Cox regression analysis of candidate genes; TPI1 exhibited the most substantial Hazard Ratio (HR), characterizing it as a preeminent risk factor. (C) Expression profiling of TPI1 across malignant versus paracancerous tissues within the TCGA-LIHC discovery cohort (upper) and GSE14520 validation cohort (lower). (D) Kaplan-Meier overall survival curves comparing patients with high and low TPI1 expression in the TCGA-LIHC (top) and GSE14520 (bottom) cohorts. (E) Relative mRNA expression levels of TPI1 in the immortalized human hepatocyte line (MIHA) and HCC cell lines (Huh7, SMMC-7721) determined by RT-qPCR. (F) Representative Western blot images (left) and densitometric quantification (right) of TPI1 protein levels in MIHA, Huh7, and SMMC-7721 cells. GAPDH served as the internal loading control. Data are expressed as mean ± SD. ** P < 0.01, *** P < 0.001.

Article Snippet: The human HCC cell lines (Huh7 and SMMC-7721) and the immortalized human hepatocyte line MIHA were procured from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Biomarker Discovery, Selection, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of BDNF treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 10 ng/mL, 50 ng/mL, or 100 ng/mL of BDNF for 48h. (A) Cells were stained for ßIII-tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count depending on BDNF concentration, and (E) neurite thickness depending on BDNF concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 images per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. BDNF: Brain-derived neurotrophic factor.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Staining, Software, Concentration Assay, Comparison, Derivative Assay

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Effect of rotenone treatment on the neurite outgrowth parameters in SH-SY5Y cells Differentiated SH-SY5Y cells were treated with 0.05 μM, 0.1 μM or 0.5 μM of rotenone for 48h. (A) Cells were stained for ß3 tubulin (red color) and DAPI (blue color). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness depending on rotenone concentration. Scale bar: 50 μm. Each data set represents N=2 independent experiments with 4-6 replicates per condition and with 5-9 pictures per well. The points represent the mean ± SEM for each condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Staining, Software, Concentration Assay, Comparison

Journal: STAR Protocols

Article Title: Protocol for automated quantification of neuronal cells using the Agilent BioTek Cytation system and Gen5 neurite outgrowth module

doi: 10.1016/j.xpro.2026.104485

Figure Lengend Snippet: Impact of the APP and P301L mutation on the neurite outgrowth capacity of SH-SY5Y cells The control condition corresponds to the native cells. The APP condition corresponds to the cells stably overexpressing the Amyloid Precursor Protein (APP). The P301L condition corresponds to the cells stably overexpressing the P301L-Tau mutation. The differentiated cells were treated with 50 ng/mL of BDNF for 48h. (A) Cells were stained for ß3 tubulin (red) and DAPI (blue). All images were captured 48h post-treatment at 20× magnification on the Agilent BioTek Cytation 5 instrument. Image analysis was performed with the Gen5 software Neurite Outgrowth module where the soma mask overlay is displayed in blue and the neurite mask overlay in yellow. (B) Average neurite length, (C) average neurite branches, (D) average neurite count, and (E) neurite thickness in Control, APP, and P301L cells. Scale bar: 50 μm. The boxes represent the median (full line) and the mean (“+” symbol); the whiskers represent the minimum and maximum values. Each data set represents N=2 independent experiments with 4-6 replicates per condition. One-way ANOVA and post hoc Dunnett’s multiple comparison tests. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The native human neuroblastoma cell line SH-SY5Y comes from ATCC (CRL-2266).

Techniques: Mutagenesis, Control, Stable Transfection, Staining, Software, Comparison

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Glycolysis inhibitors diminish the virus sensitivity of glycolytic PDAC cells MIA PaCa-2 and PK-59 cells were treated with SCH772984 (SCH) (200 nM) or 2DG (2 mM), followed by infection with OBP-401 (100 MOI) or OBP-702 (10 MOI). (A) Lactate secretion by MIA PaCa-2 and PK-45H cells treated with SCH772984 or 2DG, presented as fold-increase compared with the control group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (B) Cell lysates of MIA PaCa-2 and PK-45H cells treated with SCH or 2DG for 48 h were subjected to western blot analysis for ERK1/2, GLUT1, and LDHA. (C) MIA PaCa-2 and PK-45H cells were treated with SCH or 2DG, followed by infection with OBP-401 (100 MOI) for 24 or 48 h. Upper panels show representative photographs of immunocytochemical staining for GFP in each group 48 h after infection. Scale bars, 500 μm. Lower graphs show the fluorescence intensity of GFP analyzed under fluorescence microscopy. Data are expressed as mean (SD) of independent experiment ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D) MIA PaCa-2 and PK-45H cells were co-treated with OBP-702 and SCH772984 or 2DG at the indicated dose for 72 h. Cell viability was quantified using the XTT assay and calculated relative to the mock-infected group. Data are expressed as mean (SD) of independent experiment ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (E) Cell lysates of MIA PaCa-2 and PK-45H cells co-treated with SCH or 2DG and OBP-702 (10 MOI) for 48 h were subjected to western blot analysis for E1A, p53, PARP, and cleaved C-PARP. β-actin was assayed as a loading control. The expression level of each protein was calculated relative to that of mock-treated cells, which was set at 1.0. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Virus, Infection, Control, Western Blot, Staining, Fluorescence, Microscopy, XTT Assay, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: p53 activation modulates glutamine metabolism in PDAC cells (A) Glutamine consumption in PDAC cells, presented as fold-increase compared with PBS, which was set as 1.0. (B) Outline of glutamine metabolism, shown from glutamine uptake to α-KG production. (C) Lysates of PDAC cells were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (D) PDAC cells were infected with OBP-301 or OBP-702 at an MOI of 100 for 48 h. The amount of intracellular α-KG in PDAC cells is shown as fold-increase compared with the mock-infected group, which was set as 1.0. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (E) PDAC cells were infected with OBP-301 or OBP-702 at the indicated MOIs for 72 h. Cell lysates were subjected to western blot analysis for GDH1/2, OGDH, and IDH1. (F) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 24 h. The amount of intracellular αKG in PDAC cells is presented as fold-increase compared with mock-infected control groups. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences among four groups was determined using one-way ANOVA followed by Turkey’s multiple comparison procedure. (G) MIA PaCa-2 and PK-59 cells were infected with DL312 or Adp53 at the indicated MOIs for 48 h. Cell lysates were subjected to western blot analysis for p53, GDH1/2, OGDH, and IDH1. β-Actin was assayed as a loading control. The expression level of each protein was calculated relative to that of MIAPaCa-2 cells or mock-treated cells, which was set at 1.0. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activation Assay, Western Blot, Infection, Control, Comparison, Expressing

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Comparison of metabolic phenotypes and virus sensitivity in subcutaneous tumor models with glycolytic and non-glycolytic PDAC cells (A) Representative photographs of immunohistochemical staining for LDHA, GLUT1, and IDH1 in each group. Scale bars, 100 μm. (B) Expression levels of LDHA, GLUT1, and IDH1, calculated by dividing the DAB intensity by the number of cells in randomly selected fields. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (C) MIA PaCa-2 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (green arrows) every other day for 3 cycles. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. (D) PK-59 tumor-bearing mice received intratumoral injections of PBS (black arrows) or OBP-702 (orange arrows). The upper right photographs show tumor-bearing mice in the control and OBP-702-treated groups. The lower right photographs show tumors in the mock and OBP-702 groups. Data are expressed as mean (SD) of independent experiments ( n = 5). The statistical significance of differences between two groups was determined using the Student’s t test. ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Comparison, Virus, Immunohistochemical staining, Staining, Expressing, Control

Journal: Molecular Therapy Oncology

Article Title: Mitochondrial inhibition enhances the sensitivity of pancreatic ductal adenocarcinoma cells to oncolytic adenovirus

doi: 10.1016/j.omton.2026.201180

Figure Lengend Snippet: Investigation of the relationship between PET/CT metabolic parameters and glycolytic activity of PDAC tumors (A and B) PET/CT images of MIA PaCa-2 tumor (A) and PK-59 tumor (B). The upper left (a) shows the horizontal section, whereas the lower left (b) shows the sagittal section, and the right (c) shows the coronal section. Dotted circles indicate the tumor area. (C) Comparison of SUVmax values for MIA PaCa-2 and PK-59 tumors. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (D and E) Comparison of MTV (D) and TLG (E) values for MIA PaCa-2 and PK-59 tumors at the indicated thresholds. Data are expressed as mean (SD) of independent experiments ( n = 3). The statistical significance of differences between two groups was determined using the Student’s t test. (F and G) Scatter diagrams demonstrating correlations between expression of LDHA (F) or GLUT1 (G) and preoperative SUVmax (left), MTV (40%) (center), and TLG (40%) (right) values in patients with PDAC ( n = 30). The statistical significance of the correlations in the scatterplots was determined using Pearson’s correlation analysis. N.S., not significant; ∗, p < 0.05.

Article Snippet: Two human PDAC cell lines (MIA PaCa-2 and Capan-2) were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Positron Emission Tomography-Computed Tomography, Activity Assay, Comparison, Expressing