human bladder cancer cell lines ht1197  (ATCC)

Journal: bioRxiv

Article Title: ARID1A-mutant and deficient bladder cancer is sensitive to EZH2 pharmacologic inhibition

doi: 10.1101/2021.01.12.426383

Figure Lengend Snippet: Bladder cancer cell lines with ARID1A mutation are sensitive to EZH2 inhibition. (A) Immunoblot shows expression of ARID1A in different bladder cancer cell lines including ARID1A mutant (HT1197, HT1376, and VMCUB-1), and ARID1A wild type (T24, 5637, and RT112) cell lines. (B) Immunoblot analysis showing expression level of EZH2, Histone H3 trimethyl lysine 27 (H3K27me3) and total Histone 3 in ARID1A mutant bladder cancer cell lines (HT1197, HT1376 and VM-CUB1) after treatment with EZH2 small molecule inhibitor GSK126. (C) Cell proliferation assay indicated ARID1A mutant bladder cancer cells are sensitive to GSK126. (D) Immunoblot analysis as above in (B) in ARID1A wild type bladder cancer cell lines (T24, 5637 and RT-112) after GSK126 treatment. (E) Cell proliferation assay of ARID1A wild type bladder cancer cells showed no effect of GSK126 treatment. “ns” – non-significant.

Article Snippet: HEK293T (ATCC) and all bladder cancer cell lines HT1197 (ATCC, Manassas, VA, USA), HT1376 (ATCC), T24 (ATCC), 5637 (ATCC), RT112, VM-CUB1 (DSMZ, Braunschweig, Germany) were grown in Dulbecco’s 90% Dulbecco’s MEM (4.5 g/L glucose) with penicillin– streptomycin (100 U/ml) and 10% fetal bovine serum (Sigma-Aldrich, St Louis, MO, USA) in 5% CO 2 cell culture incubator.

Techniques: Mutagenesis, Inhibition, Western Blot, Expressing, Proliferation Assay

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: A. Mutation plots of cell lines adapted from Nickerson, M L et al. “Molecular Analysis of Urothelial Cancer Cell Lines for Modeling Tumor Biology and Drug Response.” Oncogene (2016) (27) under a Creative Commons Attribution- NonCommercial-ShareAlike4.0 International License. Cell viabilities of UMUC3, T24 and HT1197 bladder cancer cell lines treated with increasing doses of B. Cisplatin, C. Panobinostat D. Belinostat, E. Tubastatin, and F. Mocetinostat. Dose response curves showing percent viability normalized to vehicle controls after 72 hours of treatment plotted against the logarithmic drug concentrations. Half-maximal inhibitory concentrations (IC50s) were calculated using GraphPad Prism. The error bars represent biological replicates.

Article Snippet: Cell culture and Cell viabilities Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Mutagenesis

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: Significantly enriched Hallmark gene sets in HT1197 cells treated with panobinostat for 48 hours

Article Snippet: Cell culture and Cell viabilities Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques:

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: Cell cycle changes by flow cytometry using 12 nM panobinostat in A. HT1197, B. UMUC3, and C. T24 vs vehicle treatment for 24 hours. D. Changes in percent population of cells quantified by graph pad prizm. The change in the cell population distribution was replicated for a total of three times. An unpaired student t-test to compare the percent population with and without treatment showed the changes in G0/G1 and G2/M populations to be statistically significant. The error bars represent biological replicates.

Article Snippet: Cell culture and Cell viabilities Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Flow Cytometry

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: A. Western blot to assess p21 levels in bladder cancer cell lines treated with panobinostat at 2 and 4 times the IC50 dose for 48 hours. This was repeated for a total of three times B. Western blot showing accumulation of gH2Ax marks in HT1197 upon treatment with panobinostat and belinostat. This was repeated for a total of three times. C. RT PCR to assess expression of TNFα, IFNβ1 and IFNα2 in HT1197 cells treated with panobinostat and belinostat. Results from three biological replicates are represented with error bars and the change in expression is tested for significance by the unpaired t-test (Graphpad Prism).

Article Snippet: Cell culture and Cell viabilities Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: A. Mutation plots of cell lines adapted from Nickerson, M L et al. “Molecular Analysis of Urothelial Cancer Cell Lines for Modeling Tumor Biology and Drug Response.” Oncogene (2016) (27) under a Creative Commons Attribution- NonCommercial-ShareAlike4.0 International License. Cell viabilities of UMUC3, T24 and HT1197 bladder cancer cell lines treated with increasing doses of B. Cisplatin, C. Panobinostat D. Belinostat, E. Tubastatin, and F. Mocetinostat. Dose response curves showing percent viability normalized to vehicle controls after 72 hours of treatment plotted against the logarithmic drug concentrations. Half-maximal inhibitory concentrations (IC50s) were calculated using GraphPad Prism. The error bars represent biological replicates.

Article Snippet: Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Mutagenesis

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: Significantly enriched Hallmark gene sets in HT1197 cells treated with panobinostat for 48 hours

Article Snippet: Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques:

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: Cell cycle changes by flow cytometry using 12 nM panobinostat in A. HT1197, B. UMUC3, and C. T24 vs vehicle treatment for 24 hours. D. Changes in percent population of cells quantified by graph pad prizm. The change in the cell population distribution was replicated for a total of three times. An unpaired student t-test to compare the percent population with and without treatment showed the changes in G0/G1 and G2/M populations to be statistically significant. The error bars represent biological replicates.

Article Snippet: Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Flow Cytometry

Journal: Molecular cancer therapeutics

Article Title: Histone deacetylase inhibition has targeted clinical benefit in ARID1A mutated advanced urothelial carcinoma

doi: 10.1158/1535-7163.MCT-17-0957

Figure Lengend Snippet: A. Western blot to assess p21 levels in bladder cancer cell lines treated with panobinostat at 2 and 4 times the IC50 dose for 48 hours. This was repeated for a total of three times B. Western blot showing accumulation of gH2Ax marks in HT1197 upon treatment with panobinostat and belinostat. This was repeated for a total of three times. C. RT PCR to assess expression of TNFα, IFNβ1 and IFNα2 in HT1197 cells treated with panobinostat and belinostat. Results from three biological replicates are represented with error bars and the change in expression is tested for significance by the unpaired t-test (Graphpad Prism).

Article Snippet: Cell Culture Human bladder cancer cell lines HT1197, T24 and UMUC3, were obtained from ATCC between 2015 and 2018.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing