human cell line rt112  (Genentech inc)

Journal: Microbiome

Article Title: Dietary fibre supplementation enhances radiotherapy tumour control and alleviates intestinal radiation toxicity.

doi: 10.1186/s40168-024-01804-1

Figure Lengend Snippet: Fig. 5 Bacterial supernatants from the cocultures of B. acidifaciens and F. prausnitzii conferred stronger anti-tumour phenotypes in bladder cancer cells. a The five bacterial supernatants used in this experiment. b Western blot analysis of histone acetylation (N = 3) of RT112 cells treated with different bacterial supernatants. Histone acetylation levels were determined after treating with 100 mL bacterial supernatants in 2 mL of medium for 24 h. c The cell survival of RT112 cells treated with 200 mL of GAM broth or bacterial supernatants in 500 mL of medium for 2 days (N = 3). d Immunofluorescence microscopy analysis of γ-H2AX levels (N = 3) in RT112 cells treated with 100 mL bacterial supernatants in 2 mL of medium for 24 h. DNA damage was evaluated after treating with 2 Gy IR. Kruskal-Wallis test and Dunn’s multiple comparison tests were conducted to compare the means of different groups. e Linear quadratic survival curves of RT112 cells treated with 100 or 400 μL bacterial supernatant from BA+FP for 24 h before receiving irradiation of 0–8 Gy (N = 3). b, c and e One-way ANOVA with Bonferroni’s multiple comparison test was used to compare the means of different dietary groups. f Principal component analysis of known metabolites of different bacterial supernatants. The clustering effect was assessed using the ADONIS test (R2 = 0.6495, Pr(>F) = 0.001). g Relative levels of metabolites enriched in the bacterial supernatant of BA+FP. ANOVA test, followed by post-hoc analysis using Fisher’s LSD and p value adjustment using the Benjamin-Hochberg method, was applied to identify the metabolites with significantly different levels across the groups. pHs of GAM broth and bacterial supernatants were all neutralised to 7.2. BA+FP denotes the co-culture of B. acidifaciens and F. prausnitzii, while Bif+FP denotes the co-culture of Bifidobacterium and F. prausnitzii. Data are presented as mean ± SD

Article Snippet: The RT112 human bladder carcinoma cell line was obtained from DSMZ (Germany) and cultured in RPMI1640 medium (Sigma) supplemented with 10% fetal bovine serum (Invitrogen).

Techniques: Western Blot, Immunofluorescence, Microscopy, Comparison, Irradiation, Co-Culture Assay

Journal: Cell Reports Medicine

Article Title: Immunomodulatory effects and improved outcomes with cisplatin- versus carboplatin-based chemotherapy plus atezolizumab in urothelial cancer

doi: 10.1016/j.xcrm.2024.101393

Figure Lengend Snippet: The direct effects of cisplatin versus carboplatin on downstream immune-related programs in cancer cells are mediated by the DNA damage transducer ATR (A) Enrichment of Hallmark gene sets with cisplatin (red) versus carboplatin (gray) treatment in the 5637 human bladder cancer cell line (top), RT112 human bladder cancer cell line (middle), and MC38 murine colon cancer cell line (bottom). The cisplatin and carboplatin concentrations used were 5 and 35 μM, respectively, and cells were collected 24 h after treatment. Only pathways that were significantly changed with cisplatin versus carboplatin (false discovery rate [FDR] < 0.05) are shown. n = 3 per treatment group for the 5637 and RT112 cell lines; n = 2 per treatment group for the MC38 cell line. (B) Protein expression of PD-L1 as determined by the median fluorescence intensity in the three cell lines treated with increasing concentrations of cisplatin (red) or carboplatin (black) for 24 h. Data depict the aggregate of three independent experiments (mean ± SEM). (C) Representative immunoblots showing p-ATM S1981, p-ATR T1989, p-Chk1 S317, p-Chk2 T68, and GAPDH levels in lysates of the 5637 cell line treated with increasing concentrations of cisplatin or carboplatin for 6 h. One representative experiment out of four independent experiments is shown. (D) Protein expression of PD-L1 as determined by median fluorescence intensity in the 5637 cell line treated with increasing concentrations of cisplatin or carboplatin for 24 h, in the presence or absence of an ATM inhibitor (KU-55933, 1 μM) or ATR inhibitor (VE-821, 1 μM). Data depict the aggregate of three independent experiments (mean ± SEM). (E and F) Bulk RNA-seq analysis of 5637 cells treated with 5 μM cisplatin or 35 μM carboplatin, in the absence (gold) or presence of 1 μM ATR inhibitor (navy blue) for 24 h. (E) Enrichment of Hallmark gene sets. n = 4 per treatment group. (F) Heatmap displaying the scaled transcriptional expression of genes involved in immune-related transcriptional programs. Heatmap shows an average score of n = 4 individual samples per treatment group. See also Figures S5–S8 .

Article Snippet: Human cell line: RT112 , Genentech , N/A.

Techniques: Expressing, Fluorescence, Western Blot, RNA Sequencing

Journal: Cell Reports Medicine

Article Title: Immunomodulatory effects and improved outcomes with cisplatin- versus carboplatin-based chemotherapy plus atezolizumab in urothelial cancer

doi: 10.1016/j.xcrm.2024.101393

Figure Lengend Snippet:

Article Snippet: Human cell line: RT112 , Genentech , N/A.

Techniques: Blocking Assay, Recombinant, Cell Isolation, Activation Assay, Enzyme-linked Immunosorbent Assay, Staining, Software

Journal: Oncology Letters

Article Title: MKP-1 overexpression is associated with chemoresistance in bladder cancer via the MAPK pathway

doi: 10.3892/ol.2020.11741

Figure Lengend Snippet: MKP-1 and MAPK expression in RT112 cells transfected with NC and MKP-1 siRNA. (A) Relative MKP-1 expression in the siNC and siMKP-1 groups was examined using reverse transcription-quantitative PCR. **P<0.01. (B) Representative microscopic images of siNC and siMKP-1 treated cells captured in both 2D and 3D environments under a phase contrast microscope. (C) MKP-1 expression of the siNC and siMKP-1 groups, as determined via western blotting. GAPDH was used as the internal control. (D) Phosphorylated and total ERK1/2 protein expressions of the siNC and siMKP-1 group, as determined via western blotting. GAPDH was used as the internal control. (E) Phosphorylated and total p38 protein expressions of the siNC and siMKP-1 group, as determined via western blotting. GAPDH was used as the internal control. (F) Phosphorylated and total JNK protein expression levels of the siNC and siMKP-1 group, as determined via western blotting. GAPDH was used as the internal control. MKP-1, mitogen activated protein kinase phosphatase-1; NC, negative control; siMKP-1, MKP-1 small interfering RNA; siNC, small interfering negative control.

Article Snippet: The human bladder cancer cell line RT112 was purchased from the Leibniz Institute DSMZ and maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere containing of 5% CO 2 .

Techniques: Expressing, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Microscopy, Western Blot, Control, Negative Control, Small Interfering RNA