human epithelial urinary bladder cancer cell line t24  (ATCC)

Journal: Cancer research

Article Title: A urinary drug-disposing approach as an alternative to intravesical chemotherapy for treating non-muscle invasive bladder cancer

doi: 10.1158/0008-5472.CAN-21-2897

Figure Lengend Snippet: A, Bdd does not trigger any innate immune response. No increase in the inflammatory cytokines concentrations was detected in the plasma of female BALB/cJ mice (n=3/group) 4 h after i.v. administration of the Bdd peptide (5 mg/kg, 150 μL). LPS was used as a positive control and concentrations of each cytokine were measured by ELISA kit. B, Cellular uptake of Cyanine5.5-labeled Bdd (Cy-Bdd). Representative fluorescence microscopic images of human UMUC-3 BC cells and murine Renca renal adenocarcinoma cells incubated for 6 and 24 h with Cy-Bdd (0.5 nmol). Dapi (9 μM) and LysoTracker-GFP (1 μM) were used for nuclear (blue) and organelle (green) staining, respectively, and were added to the cells 30 min prior to imaging. Scale bar is 25 μm. C, Comparing the potency of different chemotherapeutics (DM1, GEM, MIT, CIS, and DOX). UMUC-3 and Renca cells were incubated with the drugs at various concentrations for 72 h prior to measuring the cell viability. The dose response curves were plotted and the half maximal inhibitory concentrations (IC50 values) of each drug calculated using Graph Pad Prism 6.0 software. D, Conjugation of DM1 to Bdd. The cleavable linker SPDP was first conjugated to the peptide N-terminal in solid phase. DM1 was then added to the cleaved peptide in a solution mixture of PBS and NMP. E, Plot showing the percentage of accumulated DM1 released from the DM1-Bdd over time in PBS in the absence and presence of GSH (1 mM). The amount of drug released was quantified using rp-HPLC analysis (absorbance detected at 254 nm). F, Conjugation of aldox to Bdd. The peptide, supplemented with a N-terminal cysteine, was incubated with aldox in PBS (pH = 7.4) for 30 min prior to purification by rp-HPLC in neutral conditions. G, Plots showing the percentage of the accumulated DOX active metabolite released from aldox-Bdd (100 μM) over time in PBS buffers with different pH values. The amount of drug released was quantified using rp-HPLC analysis (absorbance detected at 480 nm). H, DM1-Bdd displays a similar cytotoxicity compared to free drug against murine bladder (MB49), human bladder (UMUC-3 and T24), and murine kidney (Renca) cancer cell lines. Plots of relative cell viability against the drug concentration. I, Aldox-Bdd is more potent than free aldox. Plots of the relative cell viability against the drug concentration.

Article Snippet: UMUC-3 and T24 human urinary bladder cancer cell lines (Cat# CRL-1749 and Cat# HTB-4), and Renca murine kidney adenocarcinoma cell line (Cat# CRL-2947), were obtained from ATCC (Manassas, VA).

Techniques: Clinical Proteomics, Positive Control, Enzyme-linked Immunosorbent Assay, Labeling, Fluorescence, Incubation, Staining, Imaging, Software, Conjugation Assay, Purification, Concentration Assay

Journal: Molecular Oncology

Article Title: BCG‐induced cytokine release in bladder cancer cells is regulated by Ca 2+ signaling

doi: 10.1002/1878-0261.12397

Figure Lengend Snippet: BCG evokes intracellular Ca 2+ signaling in bladder cancer cells. Primary human bladder cancer cells (A) and human T24 cells (B) exposed to BCG (4 × 10 6 –6 × 10 7 cfu·mL −1 ) in a Ca 2+ ‐containing buffer exhibit Ca 2+ signaling. Primary human bladder cancer cells (C) and human T24 cells (D) exposed to BCG (4 × 10 6 –6 × 10 7 cfu·mL −1 ) in a Ca 2+ ‐free buffer also exhibit Ca 2+ signaling. (E) The number of human T24 cells responding to BCG with Ca 2+ signaling was significantly reduced by the inhibitors CPA, 2APB, U73122, ET‐18‐OCH3 (ET‐18), and PTX, whereas wortmannin failed to significantly reduce the number of active cells. Results are means ± SEM of measurements from at least three separate cell cultures. * P < 0.05, ** P < 0.01, *** P < 0.001 (Student's t ‐test).

Article Snippet: The urinary bladder cancer cell lines T24 (human), RT4 (human), and MB49 (mouse) were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA) and were occasionally tested for mycoplasma (last tested in 2015).

Techniques:

Journal: Molecular Oncology

Article Title: BCG‐induced cytokine release in bladder cancer cells is regulated by Ca 2+ signaling

doi: 10.1002/1878-0261.12397

Figure Lengend Snippet: BCG triggers cytokine release in bladder cancer cells. Primary human bladder cancer cells derived from one male tumor (A) and one female tumor (B) or mouse MB49 cells (C) exposed to BCG (4 × 10 6 –6 × 10 7 cfu·mL −1 ) secrete multiple cytokines, as compared to a control group. Results are means ± SEM of measurements from four separate cell cultures. * P < 0.05, ** P < 0.01, *** P < 0.001 (Student's t ‐test)

Article Snippet: The urinary bladder cancer cell lines T24 (human), RT4 (human), and MB49 (mouse) were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA) and were occasionally tested for mycoplasma (last tested in 2015).

Techniques: Derivative Assay, Control