human bladder transitional cancer cells  (ATCC)

Journal: The Journal of urology

Article Title: Intravenous siRNA silencing of survivin enhances activity of mitomycin C in human bladder RT4 xenografts

doi: 10.1016/j.juro.2015.02.036

Figure Lengend Snippet: RT4 cells were incubated for 4 hr in transfection medium (control), with PPCat-siNT (100 nM) or PPCat-siSurvivin (100 nM), followed by 2 hr treatment with complete medium or MMC (0.1 and 3 µM). At 48 hr post-treatment, cells were collected and analyzed for survivin mRNA and protein levels. Ctl: control. –si: no siRNA, siNT: nontarget siRNA, siSur: survivin siRNA. Data for mRNA levels are mean+1 SD (n=3 experiments, with triplicate samples per experiment). Data for protein levels are mean+1 SD (n=3 experiments, single sample per experiment). *p<0.05 compared to control. **p<0.05 compared to MMC and MMC+PPCat-siNT.

Article Snippet: Human bladder transitional RT4 cancer cells (ATCC, Manassaas, VA) were maintained in complete medium comprising Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids, 100 μg/ml penicillin and 100 μg/ml streptomycin, in 5% CO 2 at 37°C.

Techniques: Incubation, Transfection, Control

Journal: The Journal of urology

Article Title: Intravenous siRNA silencing of survivin enhances activity of mitomycin C in human bladder RT4 xenografts

doi: 10.1016/j.juro.2015.02.036

Figure Lengend Snippet: Survivin silencing enhances MMC activity in vitro and in vivo Cultured RT4 cells and mice bearing RT4 xenograft tumors were treated with MMC, nontarget siRNA and survivin siRNA (siRNA were delivered in PPCat), as described in the legends of Figures 1 and . For the in vitro studies, the MMC concentration was 3 µM and the siRNA concentration was 100 nM. For the in vivo studies, the MMC dose was 3 mg/kg given every 4 days for a total of 3 doses and the siRNA dose was 1 nmole given 2 days after each MMC treatment.

Article Snippet: Human bladder transitional RT4 cancer cells (ATCC, Manassaas, VA) were maintained in complete medium comprising Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids, 100 μg/ml penicillin and 100 μg/ml streptomycin, in 5% CO 2 at 37°C.

Techniques: Activity Assay, In Vitro, In Vivo, Cell Culture, Concentration Assay, Expressing, Control, TUNEL Assay

Journal: The Journal of urology

Article Title: Intravenous siRNA silencing of survivin enhances activity of mitomycin C in human bladder RT4 xenografts

doi: 10.1016/j.juro.2015.02.036

Figure Lengend Snippet: Antitumor activity was evaluated in immunodeficient mice bearing subcutaneous RT4 human bladder xenograft tumors. All treatments (indicated by arrows) were administered by intravenous injections. Day 0 represents the day of treatment initiation, which corresponded to 28 days post-tumor implantation. The MMC dose was 3 mg/kg, given every 4 days for a total of 3 doses. The PPCat-siSurvivin dose was 1 nmole, given 2 days after each MMC treatment. Animals were maintained for 58 days post treatment, with tumor size measurements taken every 4 days for the first 16 days and once a week afterwards. From top to bottom: control (filled diamonds, n=5), siNT (open squares, n=5), siSurvivin (filled triangles, n=5), MMC (open diamonds, n=10), MMC+siNT (crosses, n=5), MMC+siSurvivin (filled squares, n=10). *p<0.05 vs. control and single agent siNT and siSurvivin. **p<0.05 vs. all other groups.

Article Snippet: Human bladder transitional RT4 cancer cells (ATCC, Manassaas, VA) were maintained in complete medium comprising Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids, 100 μg/ml penicillin and 100 μg/ml streptomycin, in 5% CO 2 at 37°C.

Techniques: Activity Assay, Tumor Implantation, Control

Journal: The Journal of urology

Article Title: Intravenous siRNA silencing of survivin enhances activity of mitomycin C in human bladder RT4 xenografts

doi: 10.1016/j.juro.2015.02.036

Figure Lengend Snippet: RT4 cells were incubated for 4 hr in transfection medium (control), with PPCat-siNT (100 nM) or PPCat-siSurvivin (100 nM), followed by 2 hr treatment without or with MMC in complete medium, and then processed for short term MTT assay (cytotoxicity measured at 48 hr post-treatment, Panel A), or the long term clonogenic assay (cytotoxicity measured at 23 days post treatment, Panel B). For Panel A, data are mean+1 SD (n=3 experiments, with triplicate samples per experiment). Some SD values are smaller than the symbols. For Panel B, data are mean+1 SD (n=3 experiments, triplicate samples per experiment). Ctl: control. –si: no siRNA, siNT: nontarget siRNA, siSur: survivin siRNA. *p<0.05 compared to untreated control (i.e., no MMC, no siRNA). **p<0.05 compared to untreated control as well as single agent MMC at same MMC concentration (i.e., no siRNA or MMC+siNT).

Article Snippet: Human bladder transitional RT4 cancer cells (ATCC, Manassaas, VA) were maintained in complete medium comprising Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS), 1% non-essential amino acids, 100 μg/ml penicillin and 100 μg/ml streptomycin, in 5% CO 2 at 37°C.

Techniques: Incubation, Transfection, Control, MTT Assay, Clonogenic Assay, Concentration Assay