umuc3 human bladder cancer cells  (ATCC)

Journal: Translational Oncology

Article Title: A CEBPB/TYMP/GDF15 signaling axis mediates tumor growth and cisplatin resistance in bladder cancer

doi: 10.1016/j.tranon.2025.102537

Figure Lengend Snippet: Knockdown of TYMP suppresses tumor growth and EMT in BC cells. (A) Western blot detection of TYMP, E-cadherin (E-cad), N-cadherin (N-cad), and Snail in UMUC3 cells transduced with scramble control or three independent shTYMP constructs. (B–D) Cell viability (B), invasion (C), and migration (D) assays for scramble and shTYMP UMUC3 cells. (E–G) Validation of TYMP knockdown in T24 cells by CCK-8 proliferation assay (E), invasion assay (F), and migration assay (G). (H–I) TYMP knockdown in MB49 cells confirmed by western blot (H) and RT-qPCR (I). (J–L) In vivo tumor growth in C57BL/6 mice subcutaneously injected with scramble or shTymp MB49 cells; representative tumor images (J), tumor weight (K), and tumor volume (L) are shown ( n = 4 mice per group). (M) IHC staining of TYMP in xenograft tumors; scale bars: 500 μm (upper) and 50 μm (lower). (N) Immunofluorescence staining of Ki-67, N-cad, and E-cad in tumor sections, with quantification of positive area percentages. Scale bars: 50 μm. Data are presented as mean ± standard deviation (SD), and significance was determined by unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: HEK293 T cells and UMUC3 human bladder cancer cells were obtained from ATCC, while MB49 murine bladder cancer cells were sourced from Meisen CTCC (Zhejiang, China).

Techniques: Knockdown, Western Blot, Transduction, Control, Construct, Migration, Biomarker Discovery, CCK-8 Assay, Proliferation Assay, Invasion Assay, Quantitative RT-PCR, In Vivo, Injection, Immunohistochemistry, Immunofluorescence, Staining, Standard Deviation, Two Tailed Test

Journal: Translational Oncology

Article Title: A CEBPB/TYMP/GDF15 signaling axis mediates tumor growth and cisplatin resistance in bladder cancer

doi: 10.1016/j.tranon.2025.102537

Figure Lengend Snippet: CEBPB upregulates the expression of TYMP promoting BC prognosis and cisplatin resistance. (A–B) Western blot (A) and RT-qPCR (B) analysis of TYMP expression in UMUC3 cells treated with DMSO or cisplatin (10 μM). (C–D) Western blot (C) and RT-qPCR (D) analysis of TYMP expression in cisplatin-sensitive (C-S) and cisplatin-resistant (C-R) UMUC3 cells. (E) Cisplatin IC50 curves for UMUC3 cells with scramble or shTYMP knockdown. (F) Cell viability of UMUC3 cells treated with TAS-102 (20 μM) or DMSO. (G–I) In vivo tumor growth in C57BL/6 mice bearing MB49 xenografts treated with PBS, cisplatin, TAS-102, or both ( C + T ); representative tumor images (G), tumor weight (H), and tumor volume (I) are shown ( n = 5 mice per group). (J) PROMO-predicted transcription factors for TYMP. (K) RT-qPCR analysis of TYMP mRNA after CEBPB knockdown. (L–M) Schematic of TYMP promoter constructs (WT and MUT) with predicted CEBPB-binding site (L) and luciferase reporter assay showing promoter activity in the presence of CEBPB overexpression (M). (N) Cell viability of UMUC3 cells transfected with siCEBPB and/or TYMP overexpression plasmid (OE-TYMP). (O) RT-qPCR analysis of TYMP mRNA levels under cisplatin treatment with siCEBPB and/or siTYMP. (P) Western blot validation of TYMP protein in the same conditions as (O). (Q) RT-qPCR analysis of CEBPB mRNA in C-S and C-R UMUC3 cells. (R) Cisplatin IC50 curves for UMUC3 cells transfected with siCEBPB and/or OE-TYMP. (S) Correlation analysis between TYMP and CEBPB mRNA levels in TCGA BC cohort. (T) CEBPB expression in BC patients stratified by stage, invasion, and grade. (U) Kaplan–Meier analysis of overall survival in BC patients with high versus low CEBPB expression in TCGA cohort. Data are presented as mean ± SD, and significance was determined by unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: HEK293 T cells and UMUC3 human bladder cancer cells were obtained from ATCC, while MB49 murine bladder cancer cells were sourced from Meisen CTCC (Zhejiang, China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Knockdown, In Vivo, Construct, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Over Expression, Transfection, Plasmid Preparation, Biomarker Discovery, Two Tailed Test

Journal: Translational Oncology

Article Title: A CEBPB/TYMP/GDF15 signaling axis mediates tumor growth and cisplatin resistance in bladder cancer

doi: 10.1016/j.tranon.2025.102537

Figure Lengend Snippet: GDF15 mediates the regulation of TYMP in BC prognosis and cisplatin resistance. (A-B) RNA-sequencing of UMUC3 cells with three TYMP-targeting shRNAs was analyzed via GO pathway analysis (A), revealing significant gene changes in the "ECM" pathway in a heat map (B). (C-D) Western blot (C) and RT-qPCR (D) assessed GDF15 levels in UMUC3 cells with scramble or shTYMP. (E) Cell viability of UMUC3 cells with scramble or shTYMP treated with rhGDF15 (10 μM) was evaluated. (F-G) Western blot (F) and RT-qPCR (G) measured GDF15 levels in UMUC3 cells with siNC or siCEBPB. (H) Cell viability of UMUC3 cells with siNC or siCEBPB treated with rhGDF15 was assessed. (I) RT-qPCR measured GDF15 mRNA levels in C-S or C-R cells treated with cisplatin. (J-K) MB49 cells were injected into C57BL/6 mice, which received specific treatments (J), and tumor weight (K) was monitored over 24 days ( n = 5 mice per group). (L) IHC staining for Ki67 in UMUC3-derived xenograft tumors with or without cisplatin and GDF15 protein treatment. Scale bars: 500 mm (upper) and 50 mm (lower). Data are mean ± standard deviation, with significance determined by unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.001.

Article Snippet: HEK293 T cells and UMUC3 human bladder cancer cells were obtained from ATCC, while MB49 murine bladder cancer cells were sourced from Meisen CTCC (Zhejiang, China).

Techniques: RNA Sequencing, Western Blot, Quantitative RT-PCR, Injection, Immunohistochemistry, Derivative Assay, Standard Deviation, Two Tailed Test