Journal:

Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

doi: 10.1128/IAI.72.6.3171-3178.2004

Figure Lengend Snippet: Bladder cells take up AOAH precursor. (A) Uptake of pro-AOAH by T24 cells after incubation with pro-AOAH-containing medium for 5 h in the presence or absence of 10 mM M6P, G6P, or mannose. Washed cells were lysed and AOAH protein was assayed by ELISA. Each bar shows the mean and standard error of data from four independent experiments. (B) AOAH specific activity (activity/nanograms of protein) in T24 cell lysates. Compared to the pro-AOAH added in the medium (AOAH Source), cell-associated AOAH had a much greater specific activity, reflecting its activation by the T24 cells. M6P inhibited AOAH binding (A) but did not prevent the activation of the cell-associated AOAH (B).

Article Snippet: LLC-PK1 porcine proximal tubule cells (American Type Culture Collection [ATCC], CL-101), T24 human bladder cells (ATCC, HTB-4), and AOAH-transfected and untransfected BHK570 cells ( 31 ) were cultured in low-glucose Dulbecco modified Eagle medium (Invitrogen, Carlsbad, Calif.), Vitacell McCoy's 5a medium (ATCC), or DMEM with glutamine and 4.5 g of glucose (Fisher Scientific)/liter, respectively.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Activity Assay, Activation Assay, Binding Assay

Journal:

Article Title: Identification of Acyloxyacyl Hydrolase, a Lipopolysaccharide- Detoxifying Enzyme, in the Murine Urinary Tract

doi: 10.1128/IAI.72.6.3171-3178.2004

Figure Lengend Snippet: AOAH confers LPS-deacylating activity to bladder cells. T24 cells were allowed to take up AOAH for 5 h, washed, and then incubated with [3H]LPS (125 ng/ml) for the times indicated. Control cells were incubated with medium that did not contain AOAH. Whereas control and AOAH-containing cells took up similar amounts of [3H]LPS (A), only the AOAH-containing cells removed 3H-labeled fatty acids from the LPS backbone (B). The data represent combined results of three separate experiments; the error bars represent one standard error of the mean. Solid squares and bars, T24 cells with AOAH; open circles and bars, control T24 cells.

Article Snippet: LLC-PK1 porcine proximal tubule cells (American Type Culture Collection [ATCC], CL-101), T24 human bladder cells (ATCC, HTB-4), and AOAH-transfected and untransfected BHK570 cells ( 31 ) were cultured in low-glucose Dulbecco modified Eagle medium (Invitrogen, Carlsbad, Calif.), Vitacell McCoy's 5a medium (ATCC), or DMEM with glutamine and 4.5 g of glucose (Fisher Scientific)/liter, respectively.

Techniques: Activity Assay, Incubation, Control, Labeling

Journal: Cancers

Article Title: Inducing Targeted, Caspase-Independent Apoptosis with New Chimeric Proteins for Treatment of Solid Cancers.

doi: 10.3390/cancers17071179

Figure Lengend Snippet: Figure 4. GnRH-AIF chimeric protein kills target cancer cells in a dose- and time-dependent manner. Colo205 and LNCaP cells were treated with various doses of GnRH-AIF and GnRH-AIFinact (0.5, 0.7, and 1 µM) and incubated for 24, 48, or 72 h demonstrating a dose- and time-dependent cell death (A,B). Simultaneously, Colo205, LNCaP, MCF-7, SW48, Colo320, HEK293, and A549 were treated with GnRH-AIF (1 µM) and GnRH-AIFinact mutant chimera (1 µM) for 72 h and tested under the same conditions (C). A204 cells, normal fibroblasts, and T24P cancer cells lacking GnRH-R expression were treated with GnRH-AIF or PBS (D). Results (A–D) are presented as percentage of cell death calculated for treated cells as compared to control cells (treated with PBS only). All results are the average of 3–6 repeats of the experiments ± SD. p value ≤0.01 = **; p value ≤0.001 = ***; inact = inactive.

Article Snippet: A204 rhabdomyosarcoma cells (ATCC-HTB-82), T24P bladder urinary transitional cell carcinoma (ATCC-HTB-4), normal fibroblasts, (ATCC-PCS-420-013), lung A549 cells (ATCC-CCL-185), and HEK-293 renal adenocarcinoma cells (ATCC-CRL-1573) were grown in DMEM medium (01-055-1A, Sartorius) supplemented with 10% FBS, 2 mM L-glutamine, and 100 units/mL penicillin.

Techniques: Incubation, Mutagenesis, Expressing, Control

Journal: American Journal of Translational Research

Article Title: Gene expression profiles to analyze the anticancer and carcinogenic effects of arsenic in bladder cancer

doi:

Figure Lengend Snippet: The expression of DEGs belonging to different intersections. Human bladder cancer cells T24 were treated with 1 μM ATO for 1 h and 6 h. Expression values were calculated using 2-ΔΔCT method and GAPDH as the endogenous reference gene.

Article Snippet: Cell culture and treatment The T24 human bladder cancer cell line was purchased from Procell Co. Ltd., and mycoplasma testing was conducted.

Techniques: Expressing

Journal: American Journal of Translational Research

Article Title: Gene expression profiles to analyze the anticancer and carcinogenic effects of arsenic in bladder cancer

doi:

Figure Lengend Snippet: Expression of putative downstream genes of arsenic binding proteins in each expression matrix (A) or T24 cells examined by real-time PCR (B). Human T24 bladder cancer cells were treated with 1 μM ATO for 1 h and 6 h. Expression values were calculated using the 2-ΔΔCT method and GAPDH as the endogenous reference gene.

Article Snippet: Cell culture and treatment The T24 human bladder cancer cell line was purchased from Procell Co. Ltd., and mycoplasma testing was conducted.

Techniques: Expressing, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Chemotherapeutic dihydromyricetin with remarkable anti-tumor activity and biosafety for muscle invasive bladder cancer

doi: 10.3389/fphar.2025.1609354

Figure Lengend Snippet: Dihydromyricetin (DHM) inhibited the viability and proliferation of MIBC (T24 and UMUC3) cells in vitro . (a,b) To determine the appropriate DHM concentration for in vitro tests, T24 and UMUC3 cells were co-incubated with DHM at a concentration of 0, 5, 10, 20 and 30 μΜ, respectively. The relatively cell viability were assessed by MTT assay, and the value of 50% inhibiting concentration (IC50) was calculated; (c,d) The MTT proliferation curves of T24 and UMUC3 cells which were incubated with DHM at 0, 5 and 20 μM for consecutive 5 days **P < 0.01, ***P < 0.001.

Article Snippet: The muscle invasive bladder cancer (MIBC) cell lines (T24 and UMUC3) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: In Vitro, Concentration Assay, Incubation, MTT Assay

Journal: Frontiers in Pharmacology

Article Title: Chemotherapeutic dihydromyricetin with remarkable anti-tumor activity and biosafety for muscle invasive bladder cancer

doi: 10.3389/fphar.2025.1609354

Figure Lengend Snippet: Dihydromyricetin (DHM) induced MIBC (T24 and UMUC3) cells cycle arrested in G0/G1 phase. (a,b) Representative flow cytometry images of cell cycle in MIBC cells after treated with DHM for 48 h; (c,d) Quantitative results of cell cycle distribution exhibited a significantly increase in G0/G1 phase cell percentages. **P < 0.01, ***P < 0.001; (e,f) The relative expression of cell cycle related genes (P53, CDK2, CDK4, Cyclin D1, Cyclin E1) in both kinds of cells could be modulated by DHM; (g,h) Western blot images revealed an obvious downregulation of cell cycle proteins, such as CDK2, CDK4, Cyclin D1 and Ki67; (i,j) Representative immunofluorescence (IF) images of Ki67 staining. Blue signal (DAPI): cell nucleus. Red/green signal: Ki67 protein. Scale bar: 100 μm.

Article Snippet: The muscle invasive bladder cancer (MIBC) cell lines (T24 and UMUC3) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Flow Cytometry, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Chemotherapeutic dihydromyricetin with remarkable anti-tumor activity and biosafety for muscle invasive bladder cancer

doi: 10.3389/fphar.2025.1609354

Figure Lengend Snippet: Dihydromyricetin (DHM) promoted MIBC (T24 and UMUC3) cells apoptosis. (a,b) Representative flow cytometry photos of apoptosis in T24 and UMUC3 cells after treated with DHM for 48 h; (c,d) Quantitative results of cell apoptosis rate exhibited an effectively pro-apoptotic activity of DHM. *P < 0.05, **P < 0.01; (e,f) The relative expression of Caspase 3/6/9 in T24 and UMUC3 cells after DHM treatment; (g,h) Western blot images revealed a strong upregulation of apoptosis proteins.

Article Snippet: The muscle invasive bladder cancer (MIBC) cell lines (T24 and UMUC3) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Flow Cytometry, Activity Assay, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Chemotherapeutic dihydromyricetin with remarkable anti-tumor activity and biosafety for muscle invasive bladder cancer

doi: 10.3389/fphar.2025.1609354

Figure Lengend Snippet: Dihydromyricetin (DHM) inhibited the migration and survival of MIBC (T24 and UMUC3) cells in vitro . (a) The influence of DHM on MIBC cell survival was evaluated by clonogenic survival assay; (b,c) Quantitative results of cell clone number per filed. ***P < 0.001; (d) Representative images of transwell chamber assay after DHM treatment. (e,f) Transwell chamber assay indicated that the migration ability of T24 and UMUC3 cells was significantly inhibited by DHM. **P < 0.01, ***P < 0.001; (g,h) The anti-migration activity of DHM against T24 cell was investigated by wound healing assay; (i,j) The anti-migration activity of DHM against UMUC3 cell. **P < 0.01, ***P < 0.001.

Article Snippet: The muscle invasive bladder cancer (MIBC) cell lines (T24 and UMUC3) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Migration, In Vitro, Clonogenic Cell Survival Assay, Transwell Chamber Assay, Activity Assay, Wound Healing Assay

Journal: Frontiers in Pharmacology

Article Title: Chemotherapeutic dihydromyricetin with remarkable anti-tumor activity and biosafety for muscle invasive bladder cancer

doi: 10.3389/fphar.2025.1609354

Figure Lengend Snippet: Epithelial-mesenchymal transition (EMT) pathway was blocked by DHM to inhibit MIBC cells migration. (a,b) The relative gene expression of EMT bio-markers (E-cad, N-cad, Vimentin, Snail) in T24 and UMUC3 cells, respectively; (c,d) E-cad protein expression was upregulated, N-cad, Vimentin and Snail protein expression was downregulated. These results indicated that DHM could block EMT pathway, and then cause the migration inhibition of MIBC cells directly.

Article Snippet: The muscle invasive bladder cancer (MIBC) cell lines (T24 and UMUC3) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA).

Techniques: Migration, Gene Expression, Expressing, Blocking Assay, Inhibition