human blca cell lines umuc3  (ATCC)

Journal: Journal of Translational Medicine

Article Title: Glutamine metabolism reprogramming promotes bladder cancer progression via PYCR1: a multi-omics and functional validation study

doi: 10.1186/s12967-025-07386-2

Figure Lengend Snippet: Glutamine metabolism profiling and single-cell landscape in bladder cancer. ( A ) Schematic of targeted metabolomics workflow comparing BLCA tumor cells and normal urothelial cells. ( B ) Boxplot showing differential amino acid expression between tumor and normal cell lines. ( C ) UMAP clustering of single-cell transcriptomes from BLCA tissue samples. ( D ) Cell type annotation based on canonical markers. ( E ) Differential gene expression analysis (DEGs) across cell clusters. ( F ) Glutamine metabolism module score visualized by UMAP. ( G ) Urothelial cells colored by high/low glutamine score group. ( H ) Volcano plot of DEGs between high- and low-score groups. ( I ) GO enrichment analysis of DEGs showing involvement in metabolic and apoptotic pathways. Error bars indicate mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human BLCA cell lines (T24, J82, UMUC3, SW780) and a normal urothelial cell line (SV-HUC-1) were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Gene Expression

Journal: Journal of Translational Medicine

Article Title: Glutamine metabolism reprogramming promotes bladder cancer progression via PYCR1: a multi-omics and functional validation study

doi: 10.1186/s12967-025-07386-2

Figure Lengend Snippet: Knockdown or inhibition of PYCR1 suppresses proliferation, migration, and tumor growth of BLCA cells in vitro and in vivo. ( A ) Western blot showing PYCR1 expression levels in BLCA cell lines and normal urothelial cell line SV-HUC-1. ( B ) Validation of PYCR1 knockdown efficiency in T24 and J82 cells using three different shRNAs. ( C ) CCK-8 assay showing reduced proliferation of T24 and J82 cells upon PYCR1 knockdown. ( D ) Colony formation assay indicating a significant decrease in clonogenic ability after PYCR1 knockdown. ( E ) Transwell migration assay demonstrating impaired migration in PYCR1 knockdown cells, scale bar = 200 μm. ( F ) Flow cytometry analysis of cell cycle distribution in J82 and T24 cells after PYCR1 knockdown. The G2 phase showed significant accumulation with P = 0.0036 for J82 cells and P = 0.0055 for T24 cells. ( G ) Flow cytometry analysis of apoptosis in T24 and J82 cells following PYCR1 knockdown. ( H ) Schematic diagram of xenograft mouse models used to evaluate the effects of PYCR1 inhibition in vivo ( n = 6 mice per group). ( I ) Tumor volume comparison and IHC staining (H&E, PYCR1, Ki-67) of xenografts from T24 cells treated with PYCR1 inhibitor versus DMSO control. ( J ) Tumor volume comparison and IHC staining of xenografts from T24-NC and T24-shPYCR1 cells. Data are presented as mean ± SD from three independent experiments. ns indicates P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human BLCA cell lines (T24, J82, UMUC3, SW780) and a normal urothelial cell line (SV-HUC-1) were obtained from American Type Culture Collection (ATCC).

Techniques: Knockdown, Inhibition, Migration, In Vitro, In Vivo, Western Blot, Expressing, Biomarker Discovery, CCK-8 Assay, Colony Assay, Transwell Migration Assay, Flow Cytometry, Comparison, Immunohistochemistry, Control

Journal: Journal of Translational Medicine

Article Title: Glutamine metabolism reprogramming promotes bladder cancer progression via PYCR1: a multi-omics and functional validation study

doi: 10.1186/s12967-025-07386-2

Figure Lengend Snippet: PYCR1 knockdown impairs proline synthesis and suppresses the PI3K/AKT/mTOR signaling pathway in BLCA cells. ( A ) Quantification of intracellular proline levels in T24 and J82 cells after PYCR1 knockdown. ( B ) Correlation analysis showing positive associations between PYCR1 expression and SLC1A5 (left) and P5CS (right) in BLCA samples. ( C ) Western blot analysis confirming that knockdown of PYCR1 reduces protein expression levels of P5CS and SLC1A5. ( D ) Heatmap of differentially expressed genes in BLCA cells following PYCR1 knockdown from RNA-seq data. ( E - F ) Pathway enrichment analyses of downregulated ( E ) and upregulated ( F ) genes after PYCR1 knockdown, indicating involvement in PI3K-AKT and immune-related signaling pathways. ( G ) Western blot analysis validating the downregulation of PI3K, AKT, and mTOR pathway components upon PYCR1 knockdown in T24 and J82 cells. ( H ) Western blot analysis of PI3K/AKT/mTOR pathway activation following PYCR1 overexpression with or without LY294002 treatment in T24 and J82 cells. PYCR1 overexpression increased phosphorylation of PI3K, AKT, and mTOR, which was reversed by the PI3K inhibitor. Vinculin served as the loading control. Data are presented as mean ± SD from three independent experiments. * P < 0.05, ** P < 0.01; oe: Overexpression

Article Snippet: Human BLCA cell lines (T24, J82, UMUC3, SW780) and a normal urothelial cell line (SV-HUC-1) were obtained from American Type Culture Collection (ATCC).

Techniques: Knockdown, Expressing, Western Blot, RNA Sequencing, Protein-Protein interactions, Activation Assay, Over Expression, Phospho-proteomics, Control

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A Upset plot of AS events in BLCA. B Unisex Upset plot illustrating adverse events related to AS in BLCA, combined with clinical information.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques:

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A Schematic diagram showing the variation of AS events with penalty coefficient in LASSO analysis. B Schematic diagram for calculating the optimal penalty parameter λ . The x -axis represents the logarithmic ( λ ) values and the y -axis represents the degrees of freedom. The λ values selected based on the minimum criteria and the 1-SE criterion resulted in 9 AS events. C Expression trend of ACTG1 in normal and BLCA patients. D Kaplan–Meier survival curves for high and low expression groups of ACTG1. ** Indicates a significant difference between the two groups with P < 0.01, Normal (blue) = 19, Tumor (red) = 407.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A Forest plot comparing the expression of ACTG1 between BLCA and Normal groups. B Forest plot comparing the expression of ACTG1 in subgroups (Country) of BLCA and Normal groups. C and D Funnel plots for assessing publication bias and verification using trim-and-fill method. E Sensitivity analysis using leave-one-out approach; MD denotes mean difference, and 95% CI represents a 95% confidence interval.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A Tumor immune cell infiltration in BLCA patients grouped by ACTG1 expression levels. B – I Correlation of ACTG1 with 8 types of immune cells. J ssGSEA scores of BLCA patient samples classified based on low or high ACTG1 expression levels. K Expression of 20 immune checkpoint-related genes in tumors of BLCA patients grouped by ACTG1 expression levels. *Indicates statistical significance between groups, P < 0.05, **Indicates statistical significance between groups, P < 0.01. *** Indicates statistical significance between groups, P < 0.001; Low (in blue) represents the low ACTG1 expression group, High (in red) represents the high ACTG1 expression group.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A – C Volcano plots showing the DEcircRNAs in BLCA tumor tissue samples compared to normal tissue samples from three datasets: GSE92675 (tumor = 4, normal = 4), GSE97239 (tumor = 3, normal = 3), and GSE147985 (tumor = 4, normal = 4). D Intersection of DEGs in the three datasets.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques:

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: A Venn diagram showing the intersection of genes encoding DEcircRNAs and EMT-related genes. B Correspondence graph between EMT-related genes and their encoding DEcircRNAs. C – E Expression trends of EMT-related genes encoding DEcircRNAs in normal and BLCA patients; *** indicates P < 0.001 compared between the two groups; Normal (blue) = 19, Tumor (red) = 407.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques: Expressing

Journal: NPJ Precision Oncology

Article Title: ACTG1 driven splicing of P4HB gene enhances EMT and bladder cancer progression

doi: 10.1038/s41698-025-00923-8

Figure Lengend Snippet: Schematic representation of the molecular mechanism by which the variable shear regulatory factor ACTG1 regulates the splicing of EMT-related genes to generate circRNAs, promoting the growth and metastasis of BLCA.

Article Snippet: The human BLCA cell lines T24 and RT4 were cultured in McCoy’s 5a medium (catalog number: 30-2007, ATCC) containing 10% fetal bovine serum, while human T24-Luc cells and human BLCA cell line 5637 were cultured in RPMI-1640 medium (catalog number: 30-2001, ATCC) also containing 10% fetal bovine serum.

Techniques: Shear