Overexpression of CHMP2B suppresses proliferation of renal clear cell carcinoma cells

doi:10.12122/j.issn.1673-4254.2025.01.16

Abstract

Abstract:

Objective To analyze the association of CHMP2B expression level of with clinicopathological characteristics and prognosis of clear cell renal cell carcinoma (CRCC) and the possible role of CHMP2B in tumorigenesis and progression of CRCC. Methods RNAseq data of CRCC were downloaded from the TCGA database for analysis of CHMP2B expression levels in tumor and adjacent tissues and their correlation with clinicopathological characteristics of the patients. Survival outcomes of the patients with high and low CHMP2B expressions were analyzed using the Kaplan-Meier model, and the COX risk regression model was used for identifying the prognostic factors of the patients. The correlation between CHMP2B expression and immune infiltration, its co-expressed genes, and the effect of CHMP2B gene mutations on immunotherapy responses, and its immunohistochemical expression in CRCC and normal tissues were analyzed. Clinical samples of CRCC were collected to examine CHMP2B expressions using RT-PCR, and cell experiment was carried out to test the effect of CHMP2B overexpression on biological behaviors of CRCC cells. Results CHMP2B was significantly under-expressed in renal cancer tissues, and its overexpression obviously inhibited the proliferation of CRCC cells in vitro. CHMP2B expression level was significantly correlated with age, gender, lymph node metastasis, and tumor stage, and the patients with low CHMP2B expression had poor survival outcomes. Enrichment and co-expression gene analyses suggested that CHMP2B was mainly involved in viral outgrowth, necrotic apoptosis, endocytosis, and immune-regulatory processes in kidney cancer. Conclusion CHMP2B is lowly expressed in renal cancer tissues to affect tumor progression and tumor immune processes, and may serve as a prognostic biomarker and therapeutic target for CRCC.

Key words: renal clear cell carcinoma, charged multivesicular body protein 2B, endosomal sorting complex required for transport, prognosis, immunity

Xiaorui CHEN, Qingzheng WEI, Zongliang ZHANG, Jiangshui YUAN, Weiqing SONG. Overexpression of CHMP2B suppresses proliferation of renal clear cell carcinoma cells[J]. Journal of Southern Medical University, 2025, 45(1): 126-136.

Figures/Tables 12

Fig.1 Differential expression of CHMP2B in KIRC. A: Unpaired difference analysis. B: Paired difference analysis.

Fig.2 CHMP2B survival analysis in CRCC patients. A: CHMP2B ROC curve for predicting overall survival (The x axis is the false positive rate, and the x axis the true positive rate). B: Effect of high and low CHMP2B expression on overall survival of KIRC patients. C: Effect of high and low CHMP2B expression on disease-specific survival of KIRC.

Fig.3 Correlation analysis between CHMP2B expression and clinical characteristics of CRCC patients. A: Relationship between CHMP2B expression and T stage. B: Relationship between CHMP2B expression and N stage. C: Relationship between CHMP2B expression and M stage. D: Relationship between CHMP2B expression and grade. E: Relationship between CHMP2B expression and gender. F: Relationship between CHMP2B expression and age. ***P<0.001.

Fig.4 COX regression analysis. A: Univariate analysis. B: Multivariate analysis. C: Nomogram of survival rate.

Fig.5 CHMP2B was correlated with ESCRT family analysis, GO analysis and KEGG enrichment analysis in KIRC.. A: Relationship between CHMP2B and ESCRT family genes. B: GO analysis of CHMP2B. C: KEGG analysis of CHMP2B.

Fig.6 GSVA enrichment analysis. A: KEGG analysis of CHMP2B. B: GO analysis of CHMP2B.

Fig.7 Differential expression of CHMP2B in pan-cancer. *P<0.05, **P<0.01, ***P<0.001.

Fig.8 Correlation analysis between CHMP2B and tumor microenvironment. A: CHMP2B immune score. B: Relationship between CHMP2B expression and immune cells. C: Differential expression of CHMP2B in immune cells. *P<0.05, **P<0.01, ***P<0.001.

Fig.9 CHMP2B immune correlation analysis. A: Heatmap of related immune checkpoint. B: Scatter plot of immune mutation. C: Relationship between high and low CHMP2B expression and treatment with ctla 4 and pd 1.

Fig.10 CHMP2B immunohistochemistry. A: Renal clear cell carcinoma tissue. B: Normal kidney tissue.

Fig.11 Differential CHMP2B expression analyzed using RT-PCR. ****P<0.0001 vs NC.

Fig.12 Effects of CHMP2B on the biological behaviors of clear cell carcinoma cells 786-O. A: Expression level of CHMP2B gene in the transfected cells detected with real-time PCR. B: Change of cell proliferation rate assessed with CCK-8 experiment. C: Effect of overexpression of CHMP2B on horizontal migration ability of the cells (Original magnification: ×100). D: Effect of overexpression CHMP2B on vertical migration ability of cells in Transwell experiment (crystal violet staining, ×100). ****P<0.0001 vs Vector-NC.

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