Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Jun 24:15:1564826.
doi: 10.3389/fonc.2025.1564826. eCollection 2025.

CHMP6 as a novel prognostic biomarker in bladder cancer: insights from a comprehensive cell death-related gene risk model

Wei Ning #  1 , Yan-Ru Yang #  2 , Chen-Xu Wu  3 , Yue Hou  1 , Zhi-Wei Liu  1 , Chang-Bin Yang  1
Affiliations

CHMP6 as a novel prognostic biomarker in bladder cancer: insights from a comprehensive cell death-related gene risk model

Wei Ning et al. Front Oncol. .

Abstract

Introduction: Bladder cancer (BLCA) is a prevalent and aggressive disease characterized by substantial molecular heterogeneity, complicating its diagnosis and treatment. Existing therapies, including surgery and chemotherapy, often lack specificity. Alterations in cell death mechanisms, such as ferroptosis, cuproptosis, and immunogenic cell death, significantly impact cancer progression and prognosis.

Methods: We analyzed gene expression data from TCGA and GEO. Cox regression analyses generated a prognostic risk score model incorporating LIPT1, ACSL5, and CHMP6. This model successfully stratified BLCA patients into different risk categories and was validated through survival analysis, immune infiltration, mutation burden assessment, drug sensitivity predictions, and single-cell analysis. The high-risk group was linked to differentiation processes, developmental stages, and active metabolic pathways.

Results: Experimental validation highlighted CHMP6's role in enhancing BLCA cell survival and migration by regulating the cell cycle. The model's prognostic relevance was further supported by drug sensitivity and immune metrics. These results provide valuable insights into potential biomarkers and therapeutic targets for BLCA treatment.

Discussion: The CHMP6 protein promotes BLCA cell survival and invasive migration through modulation of the cell cycle.

Keywords: BLCA; CDRI; CHMP6; ICD; ferroptosis.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Differential expression and prognostic significance of genes in bladder cancer. (A) Volcano Plot of Differentially Expressed Genes between bladder cancer and normal samples. (B) Heatmap of Ferroptosis and Immunogenic Cell Death-Related Genes. (C) Forest Plot of Univariate Cox Analysis: Hazard ratios and 95% confidence intervals for genes significantly associated with bladder cancer prognosis from univariate Cox analysis. (D) Forest Plot of Multivariate Cox Analysis.
Figure 2
Figure 2
Survival analysis of risk groups (A) Overall Survival in TCGA-BLCA: Kaplan-Meier survival curves for high-risk and low-risk groups in bladder cancer (TCGA-BLCA). (B-E) Kaplan-Meier survival curves for GSE31684, TCGA-LIHC, TCGA-MESO, TCGA-UCEC, and TCGA-ACC.
Figure 3
Figure 3
Functional enrichment analysis of differentially expressed genes in bladder cancer. (A) GO Pathway Enrichment Analysis: Enriched Gene Ontology (GO) terms for differentially expressed genes include biological processes (BP), cellular components (CC), and molecular functions (MF). (B) KEGG Pathway Enrichment Analysis.
Figure 4
Figure 4
Immune infiltration and tumor microenvironment analysis. (A) Relative Abundance of Immune Cell Types in High and Low Risk Groups. (B) Comparison of Immune Cell Composition Between High and Low Risk Groups. Statistical significance is indicated by ns (not significant), * (adjust.p< 0.05), ** (adjust.p< 0.01), *** (adjust.p< 0.001), and **** (adjust.p< 0.0001). (C) Correlation Heatmap of Immune Cell Types. (D) ESTIMATE Scores Comparison Between High and Low Risk Groups.
Figure 5
Figure 5
Mutation analysis in risk groups. (A, B) Mutation Summary in Risk Group. Variant classification and type distribution. (C, D) Mutation Landscape in Risk Group. (E, F) Tumor Mutation Burden (TMB) in Risk Group.
Figure 6
Figure 6
Drug sensitivity analysis in high and low risk groups. (A) Predicted Drug Sensitivity. Boxplots compare predicted drug sensitivity between high risk and low risk groups. Statistical significance is indicated by ** (adjust.p< 0.01) and *** (adjust.p< 0.001). (B) Correlation between Drug Sensitivity and Risk Score.
Figure 7
Figure 7
Survival analysis and immune cell impact in risk groups. (A, B) Overall Survival Based on Tumor Mutation Burden (TMB) in risk Group. (C, D) Overall Survival Based on CXCR5+CD8+ T Cells in risk Group. (E) Comparing TIDE scores between the risk groups. (F) Comparing TILs between the risk groups.
Figure 8
Figure 8
Single-Cell RNA sequencing analysis of bladder cancer samples. (A) UMAP Plot of cell types. (B) Dot plot of cell types. (C) Heatmap of expression of three genes in cell types. (D) Heatmap of expression of three genes in cell phase. (E) The expression distribution of three genes in the trajectory of cell differentiation.
Figure 9
Figure 9
Validation of CHMP6 through in vitro methods. (A-C) Immunohistochemical Staining: Immunohistochemical analysis of CHMP6, ACSL5, and LIPT1 in BLCA tissues. (D) Protein Expression Levels: Expression of CHMP6 and ACSL5 proteins in 5637 and T24 cell lines. (E) Western Blot (WB) Analysis: Detection of CHMP6 protein knockdown in 5637 and T24 cell lines, along with ACSL5 expression. (F) RNA Knockdown Efficiency: Validation of CHMP6 knockdown efficiency at the RNA level in 5637 and T24 cell lines. (G) Apoptosis Assay: Assessment of early and late apoptosis in 5637-shControl and 5637-shCHMP6#2 cells. (H) Cell Cycle Analysis: Flow cytometry analysis of the cell cycle in 5637-shControl and 5637-shCHMP6#2 cells. (I) Migration Assay: Scratch assay to evaluate the migration ability of 5637-shControl and 5637-shCHMP6#2 cells. (J) Invasion and Migration Assay: Transwell assay to assess invasion and migration capabilities of 5637-shControl and 5637-shCHMP6#2 cells. ns (not significant) and ** (adjust.p < 0.01).
See this image and copyright information in PMC

References

    1. Antoni S, Ferlay J, Soerjomataram I, Znaor A, Jemal A, Bray F. Bladder cancer incidence and mortality: A global overview and recent trends. Eur Urol. (2017) 71:96–108. doi: 10.1016/j.eururo.2016.06.010 - DOI - PubMed
    1. Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, Rebelo M, et al. GLOBOCAN 2012 v1.0, cancer incidence and mortality worldwide: IARC cancer base no. 11 [Internet. Int Agency Res Cancer Lyon. (2014).
    1. Compérat E, Amin MB, Cathomas R, Choudhury A, Santis M, Kamat A, et al. Current best practice for bladder cancer: a narrative review of diagnostics and treatments. Lancet. (2022) 400:1712–21. doi: 10.1016/S0140-6736(22)01188-6 - DOI - PubMed
    1. Alifrangis C, McGovern U, Freeman A, Powles T, Linch M. Molecular and histopathology directed therapy for advanced bladder cancer. Nat Rev Urol. (2019) 16:465–83. doi: 10.1038/s41585-019-0208-0 - DOI - PubMed
    1. Caffo O, Veccia A, Fellin G, Russo L, Mussari S, Galligioni E. Trimodality treatment in the conservative management of infiltrating bladder cancer: a critical review of the literature. Crit Rev Oncol Hematol. (2013) 86:176–90. doi: 10.1016/j.critrevonc.2012.09.011 - DOI - PubMed

LinkOut - more resources