human bladder cancer cell lines t24  (Procell Inc)

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Betulinic acid (BA) enhances tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity in T24 human bladder cancer cells. T24 cells were treated with the indicated concentrations of BA and TRAIL alone or in combination for 24 h. (A, B) After treatment, cell viability was assessed via 3′-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. (C) Morphological changes in the cells treated with BA and TRAIL alone or in combination were observed under an inverted microscope. (D, E) After 4′,6-diamidino-2-phenylindole (DAPI) staining, nuclear morphological changes were observed under a fluorescence microscope (D), and the frequency of cells with chromatin condensation and fragmentation was determined (E). (F, G) Flow cytometry was performed after propidium iodide (PI) staining. Representative histograms (F) and the frequency of cells in the sub-G1 phase (G) are shown. * p <0.05, ** p <0.01, and *** p <0.001 vs. control cells; ### p <0.001 vs. BA-treated cells (n=3).

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: MTT Assay, Inverted Microscopy, Staining, Fluorescence, Microscopy, Flow Cytometry, Control

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Combination treatment with BA and TRAIL increases reactive oxygen species (ROS) production and decreases ATP levels in T24 human bladder cancer cells. T24 cells were treated with BA and TRAIL alone or in combination and cultured for 30 min (A-C) or 24 h (D). (A, B) After treatment, the cells were stained with 2′,7′-dichlorofluorescein diacetate (DCF-DA) and analyzed via flow cytometry. Representative histograms (A) and the frequency of DCF-positive cells (B) are shown. (C) Representative fluorescence images of the cells stained with DCF-DA, indicating ROS production, were captured via fluorescence microscopy. (D) Intracellular ATP levels were measured using an ATP assay kit. * p <0.05 and ** p <0.01 vs. control cells; ### p <0.001 vs. BA-treated cells (n=3).

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Cell Culture, Staining, Flow Cytometry, Fluorescence, Microscopy, ATP Assay, Control

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Combination treatment with BA and TRAIL increases mitochondrial damage and induces changes in the expression levels of Bcl-2 family proteins in T24 human bladder cancer cells. T24 cells were treated with BA and TRAIL alone or in combination and cultured for 24 h. (A, B) After treatment, the cells were stained with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetramethylbenzimidazolyl carbocyanine iodide (JC-1) and analyzed via flow cytometry. Representative histograms (A) and the frequency of cells with JC-1 monomers (B), indicating mitochondrial membrane potential (MMP) loss, are shown. * p <0.05 vs. control cells; ### p <0.001 vs. BA-treated cells (n=3). (C, D) Mitochondrial and cytosolic fractions were isolated from the cells (C), total protein was extracted (D), and immunoblotting was performed using antibodies against target proteins. Cytochrome c oxidase (COX IV) and β-actin were used as loading controls for the two fractions, respectively.

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Expressing, Cell Culture, Staining, Flow Cytometry, Membrane, Control, Isolation, Western Blot

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Combination treatment with BA and TRAIL activates the caspase-dependent extrinsic and intrinsic apoptotic pathways in T24 human bladder cancer cells. T24 cells were either directly co-treated with BA and TRAIL or pretreated with z-VAD-fmk for 1 h before co-treatment with BA and TRAIL and cultured for 24 h. (A) Total protein was extracted, and immunoblotting analysis was performed using antibodies against target proteins. (B) Caspase activity was examined using caspase assay kits. (C, D) After DAPI staining, nuclear morphological changes were observed under a fluorescence microscope (C), and the frequency of cells with chromatin condensation and fragmentation was determined (D). (E, F) Flow cytometry was performed after PI staining. Representative histograms (E) and the frequency of cells in the sub-G1 phase (F) are shown. ** p <0.01 and *** p <0.001 vs. control cells; ### p <0.001 vs. BA and TRAIL co-treated cells (n=3).

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Cell Culture, Western Blot, Activity Assay, Caspase Assay, Staining, Fluorescence, Microscopy, Flow Cytometry, Control

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Combination treatment with BA and TRAIL inhibits activation of the phosphoinositide 3-kinase (PI3K)/Akt pathway in T24 human bladder cancer cells. T24 cells were either directly co-treated with BA and TRAIL or pretreated with LY294002 for 1 h before co-treatment with BA and TRAIL and cultured for 24 h. (A) Total cellular proteins were isolated from the cells, and phosphorylation levels of PI3K and Akt were determined via immunoblotting. (B, C) After DAPI staining, nuclear morphological changes were observed under a fluorescence microscope (B), and the frequency of cells with chromatin condensation and fragmentation was determined (C). (D, E) Flow cytometry was performed after PI staining. Representative histograms (D) and the frequency of cells in the sub-G1 phase (E) are shown. (F) Cell viability was assessed via MTT assay. *** p <0.001 vs. control cells; ### p <0.001 vs. BA and TRAIL co-treated cells (n=3).

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Activation Assay, Cell Culture, Isolation, Phospho-proteomics, Western Blot, Staining, Fluorescence, Microscopy, Flow Cytometry, MTT Assay, Control

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Combination treatment with BA and TRAIL increases apoptosis in T24 human bladder cancer cells in an ROS-dependent manner. T24 cells were pretreated with N-acetyl-l-cysteine (NAC) 1 h before combination treatment with BA and TRAIL and cultured for 24 h. (A, B) Total protein was extracted and analyzed via immunoblotting using antibodies against target proteins. β-actin was used as a loading control. (C) Caspase-3 activity was measured using a caspase-3 assay kit. (D, E) After DAPI staining, nuclear morphological changes were observed under a fluorescence microscope (D), and the frequency of cells with chromatin condensation and fragmentation was determined (E). (F, G) Flow cytometry was performed after PI staining. Representative histograms (F) and the frequency of cells in the sub-G1 phase (G) are shown. (H) Cell viability was assessed via MTT assay. *** p <0.001 vs. control cells; ### p <0.001 vs. BA and TRAIL co-treated cells (n=3).

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Cell Culture, Western Blot, Control, Activity Assay, Caspase-3 Assay, Staining, Fluorescence, Microscopy, Flow Cytometry, MTT Assay

Journal: Biomolecules & Therapeutics

Article Title: Combination Therapy with Betulinic Acid and TRAIL Increases ROS-Dependent Cytotoxicity and Inhibits PI3K/Akt Signaling in Human Bladder Cancer Cells

doi: 10.4062/biomolther.2026.035

Figure Lengend Snippet: Schematic diagram of cytotoxicity induction in betulinic acid and TRAIL co-treated T24 human bladder cancer cells. TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; PI3K, phosphoinositide 3-kinase; NAC, N-acetyl-l-cysteine; MMP, mitochondrial membrane potential; FADD, Fas-associated death domain; tBid, truncation of BH3-interacting domain death agonist; PARP, poly(ADP-ribose) polymerase.

Article Snippet: T24 human bladder cancer cells obtained from the American Type Culture Collection (Manassas, VA, USA) were cultured as previously described ( Kim et al. , 2021 ).

Techniques: Membrane

Journal: Frontiers in Immunology

Article Title: Ribosome biogenesis programs define a three-gene RBscore with prognostic relevance in bladder cancer

doi: 10.3389/fimmu.2026.1810132

Figure Lengend Snippet: Experimental validation of RBscore signature gene expression. (A) RT-qPCR of PIN4, POP4, and PRKDC mRNA in SV-HUC-1 and T24 cells. (B) RT-qPCR of PIN4, POP4, and PRKDC mRNA in paired BLCA and adjacent normal tissues. (C) Western blot analysis and quantification of PIN4, POP4, and PRKDC protein in SV-HUC-1 and T24 cells. (D) Representative IHC images of PRKDC in BLCA and adjacent normal tissues. (E) Quantification of PRKDC IHC scores in BLCA and adjacent normal tissues. * P < 0.05, *** P < 0.001. BLCA, bladder cancer; RBscore, ribosome biogenesis related score; IHC, immunohistochemistry.

Article Snippet: Human bladder urothelial SV-HUC-1 cells (Procell, China) and bladder cancer T24 cells (Procell, China) were cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, USA) and 1% penicillin–streptomycin at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Biomarker Discovery, Gene Expression, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: BMC Cancer

Article Title: The potential antitumor effects of combining intravesical therapy with recombinant bacillus calmette-guérin and an anti-PD-1 inhibitor in bladder cancer

doi: 10.1186/s12885-026-15890-x

Figure Lengend Snippet: Cell viability of 3D bioprinted bladder cancer-on-a-chip model. A Live/dead staining on the 3rd day after drug treatment. T24 bladder cancer cell blocks in the rBCG- dltA and pembrolizumab combination group had lower cell survival rates than those in the other groups did. Noncancerous cell lines, such as HUVECs and MRC-5 cells, survived well in all groups. The scale bar at the lower right corner represents 200 μm. B According to the results of the quantitative cell density test, T24 cancer cells were affected by rBCG- dltA and pembrolizumab. The group with the lowest cell density was the combination-treated group. * p < 0.05 vs. control

Article Snippet: The 3D bioprinted BCOC blocks consisted of the T24 bladder cancer cells, MRC-5 fibroblasts, Jurkat T lymphocytes, THP-1 monocytes, and human umbilical vein endothelial cell line cells (HUVEC), cultured using standard protocols provided by the suppliers: Korean Cell Line Bank (Seoul, Republic of Korea) and Lonza (Basel, Switzerland).

Techniques: Staining, Control