human bladder cancer cell lines j82  (DSMZ)

Journal: Bladder Cancer

Article Title: Impact of DNA repair deficiency on sensitivity to antibody-drug conjugate (ADC) payloads in bladder cancer *

doi: 10.1177/23523735251317865

Figure Lengend Snippet: Homologous recombination (HR) deficiency confers cisplatin and PARP inhibitor sensitivity in bladder cancer cells. (a) Immunoblot showing siRNA-mediated depletion of BRCA2 protein in KU19-19 and J82 bladder cancer cell lines. Vinculin is shown as a loading control. (b) Immunofluorescence (IF) microscopy shows loss of radiation-induced Rad51 foci formation in BRCA2-depleted compared to BRCA2-intact bladder cancer cells. (c) Cell viability assays demonstrate increased sensitivity to cisplatin following BRCA2 depletion in bladder cancer cell lines. (d) Cell viability assays demonstrate increased sensitivity to PARP inhibitors olaparib and talazoparib in bladder cancer cell lines. kDa, kilodalton. NTC, non-targeting control. Gy, Gray. MFI, mean fluorescence intensity. *p < 0.01 (IC50 values). Error bars represent standard deviation of data collected from assays performed in triplicate (panel B) or quadruplicate (panels C, D).

Article Snippet: Human bladder cancer cell lines J82 and KU19-19 were purchased from ATCC and DSMZ, respectively.

Techniques: Homologous Recombination, Western Blot, Control, Immunofluorescence, Microscopy, Fluorescence, Standard Deviation

Journal: Bladder Cancer

Article Title: Impact of DNA repair deficiency on sensitivity to antibody-drug conjugate (ADC) payloads in bladder cancer *

doi: 10.1177/23523735251317865

Figure Lengend Snippet: Combined activity of MMAE and SN-38 with DNA repair inhibitors. (a) Cytotoxic activity of combining MMAE or SN-38 with ATR inhibition (berzosertib, BERZ), USP1 inhibition (ML323), or PARP inhibition (Olaparib, OLA; talazoparib, TALA) in KU19-19 (top graph) and J82 (bottom graph) bladder cancer cell lines with versus without HR deficiency conferred by BRCA2 depletion. (b) Cytotoxic activity of combining MMAE or SN-38 with ATR inhibition (berzosertib, BERZ), USP1 inhibition (ML323), or PARP inhibition (Olaparib, OLA; talazoparib, TALA) in NER-proficient ( ERCC2 WT) KU19-19 cells versus NER-deficient ( ERCC2 -mutant) KE182 cells (top graph) or NER-proficient (ERCC4 WT) H460 cells versus NER-deficient (ERCC4-deleted) H460 cells (bottom graph). Combination activity is quantified by the combination index (CI, see Methods) with positive log10(CI) values indicative of antagonism and negative log10(CI) values indicative of synergism. HRP, homologous recombination proficient (WT BRCA2); HRD, homologous recombination deficient (BRCA2 depleted); NERP, nucleotide excision repair proficient; NERD, nucleotide excision repair deficient. Error bars represent standard deviation of data collected from assays performed in quadruplicate.

Article Snippet: Human bladder cancer cell lines J82 and KU19-19 were purchased from ATCC and DSMZ, respectively.

Techniques: Activity Assay, Inhibition, Mutagenesis, Homologous Recombination, Standard Deviation

Journal: Pharmaceuticals

Article Title: Trichostatin C Synergistically Interacts with DNMT Inhibitor to Induce Antineoplastic Effect via Inhibition of Axl in Bladder and Lung Cancer Cells

doi: 10.3390/ph17040425

Figure Lengend Snippet: Characterization of TSC of its cytotoxicity and HDAC inhibitory activity. Structure of TSC (TSC) isolated from fermentation of Streptomyces sp. CPCC 203,909 ( a ). Concentration–effect curves of TSC in A549, SK-BR-3, and J82 ( b ). Acetylation levels of α-Tubulin and Histone H3. J82 cells were treated with the indicated concentrations of trichostatin A, Entinostat, and TSC ( c ).

Article Snippet: The human urothelial bladder cancer cell line J82 was obtained from Merck Millipore (Burlington, MA, USA).

Techniques: Activity Assay, Isolation, Concentration Assay

Journal: Pharmaceuticals

Article Title: Trichostatin C Synergistically Interacts with DNMT Inhibitor to Induce Antineoplastic Effect via Inhibition of Axl in Bladder and Lung Cancer Cells

doi: 10.3390/ph17040425

Figure Lengend Snippet: TSC-induced activation of caspase 3/7. J82 cells were treated with TSC with TSC for 48 h and labeled with Hoechst 33,342 for cell nucleus and CellEvent caspase 3/7 reagent for activated caspase 3/7 ( a ). Percentage of caspase 3/7-positive cells ( b ). The two-tailed Student’s t -test or ANOVA was applied. Results are presented as mean ± SEM from three experiments. * p < 0.05 compared to control cells.

Article Snippet: The human urothelial bladder cancer cell line J82 was obtained from Merck Millipore (Burlington, MA, USA).

Techniques: Activation Assay, Labeling, Two Tailed Test

Journal: Pharmaceuticals

Article Title: Trichostatin C Synergistically Interacts with DNMT Inhibitor to Induce Antineoplastic Effect via Inhibition of Axl in Bladder and Lung Cancer Cells

doi: 10.3390/ph17040425

Figure Lengend Snippet: Synergistic anti-cancer effect of TSC and decitabine in human urothelial bladder cancer cell line J82 and human lung cancer cell line A549. Total of 48 h incubation with decitabine prior to TSC significantly increased cytotoxicity of TSC in J82 ( a ) and in A549 ( b ) cell lines. Synergistic effect was determined by the Chou–Talalay method in J82 ( c ) and A549 ( d ) cell lines. Left panel and middle panels: cell survival rate upon treatment with TSC and decitabine alone or in combination. The right panel indicates the fraction of cells affected (Fa) and CI. Synergistic effect is defined by CI < 1. Results are presented as mean ± SD from three experiments. ** p < 0.01, * p < 0.05, combination treatment vs. single treatment.

Article Snippet: The human urothelial bladder cancer cell line J82 was obtained from Merck Millipore (Burlington, MA, USA).

Techniques: Incubation

Journal: Pharmaceuticals

Article Title: Trichostatin C Synergistically Interacts with DNMT Inhibitor to Induce Antineoplastic Effect via Inhibition of Axl in Bladder and Lung Cancer Cells

doi: 10.3390/ph17040425

Figure Lengend Snippet: TSC altered expression of Axl and targets involved in FoxO1 pathway. Representative Western blot analysis of J82 cells upon the treatment of TSC for 48 h ( a ). Densitometrically measure expression of proteins relative to GAPDH are shown with three independent experiments ( b ). Results shown are the mean ± SD from three independent experiments. ** p < 0.01, * p < 0.05 as compared with control cells.

Article Snippet: The human urothelial bladder cancer cell line J82 was obtained from Merck Millipore (Burlington, MA, USA).

Techniques: Expressing, Western Blot