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GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma

  • Authors:
    • Xiaobao Cheng
    • Shujiang Ye
    • Xiang Xu
    • Shuo Yang
    • Wei Zhao
    • Zhenquan Lu
    • Caiyong Lai
  • View Affiliations / Copyright

    Affiliations: Department of Urology, The Sixth Affiliated Hospital of Jinan University, Dongguan, Guangdong 523560, P.R. China, Department of Urology, The University of Hong Kong‑Shenzhen Hospital, Shenzhen, Guangdong 518053, P.R. China, Department of Orthopedics, Tongzhou People's Hospital, Nantong, Jiangsu 226334, P.R. China
    Copyright: © Cheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 571
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    Published online on: October 3, 2025
       https://doi.org/10.3892/ol.2025.15317
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Abstract

Treatment of advanced clear cell renal cell carcinoma (ccRCC) primarily involves targeted therapy and immunotherapy, however a number of patients develop treatment resistance. Identifying preferred biomarkers and immune‑related therapeutic targets is clinically important for the effective treatment of metastatic ccRCC. Due to the unmet clinical need for novel therapeutic approaches, it is important to explore potential biomarkers and their roles in tumor progression. Although guanylate‑binding protein 5 (GBP5) expression is notably upregulated in several cancers and promotes tumor progression, its role in ccRCC remains largely unknown. In the present study, GBP5 expression levels were compared between tumors and adjacent normal tissue using data from the Gene Expression Omnibus, Gene Expression Profiling Interactive Analysis 2 and Tumor Immune Estimation Resource (TIMER) databases. The relationship between GBP5 expression and clinicopathological features, along with its diagnostic and prognostic values, was evaluated using R software and the Kaplan‑Meier plotter. Gene Ontology term analysis and gene set enrichment analysis were performed to explore the biological functions of GBP5. The GeneMANIA database was used to investigate the protein‑protein interaction networks involving GBP5. The Tumor Immune Single‑Cell Hub and TIMER databases were used to evaluate the immune infiltration landscape of ccRCC and the association between GBP5 and immune cell markers. In vitro, GBP5 levels were manipulated in renal cancer cells through silencing or overexpressing GBP5 using lentivirus transfection experiments. The migratory and invasive capabilities of the cells were evaluated using wound‑healing and Transwell assays. The GBP5 levels in ccRCC tissues were found to be higher compared with those in adjacent tissues. GBP5 expression was associated with the tumor‑lymph node‑metastasis classification and pathological stage. Furthermore, elevated GBP5 level was associated with a poor prognosis and demonstrated promising diagnostic value. Bioinformatics analysis of GBP5 suggested its involvement in multiple biological processes and its notable association with tumor‑infiltrating immune cells. Cellular experiments demonstrated that GBP5 regulates the migration and invasion of renal cancer cells. Overall, GBP5 shows potential as a prognostic indicator and a candidate target for immunotherapy in patients with ccRCC.
View Figures

Figure 1

Overall workflow of the study.
Schematic representation of the major steps and analytical methods
employed, offering a concise and visual summary of the entire
research process. KIRC datasets from TCGA and GEO databases were
utilized to perform differential gene expression and prognostic
analyses of GBP1-7 genes, with a focus on the clinical pathological
characteristics of GBP5 in ccRCC, as well as the gene function
enrichment analysis, PPI network analysis, GSEA analysis and immune
cell infiltration analysis conducted. Finally, wound-healing and
Transwell assays were conducted to evaluate the role of GBP5 in the
invasion and migration of ccRCC cells. KIRC, kidney renal clear
cell carcinoma; TCGA, The Cancer Genome Atlas; GEO, Gene Expression
Omnibus; GBP, guanylate-binding protein; PPI, protein-protein
interaction; GSEA, gene set enrichment analysis; ccRCC, clear cell
renal cell carcinoma.

Figure 2

Expression and prognostic analysis of
GBP1-7 in ccRCC. (A-G) The expression levels of GBP1-7 in KIRC and
matching adjacent tissues were assessed using the gene expression
profiling interactive analysis 2 online tool. Results indicate that
GBP1, GBP2, GBP4 and GBP5 are highly expressed in KIRC tissues. The
mRNA expression of GBP1-6 in ccRCC and adjacent tissues from Gene
Expression Omnibus databases (H) GSE53757 (n=72) and (I) GSE36895
(n=23) was analyzed. Results show that GBP1-6 are highly expressed
in ccRCC tissues. (J-P) Kaplan-Meier survival analysis results
indicate that elevated levels of GBP2 and GBP5 are associated with
a poor prognosis, whereas low expression of GBP3, GBP4 and GBP7 is
associated with unfavorable outcomes. *P<0.05, **P<0.01 and
***P<0.001. GBP, guanylate-binding protein; ccRCC, clear cell
renal cell carcinoma; KIRC, kidney renal clear cell carcinoma; TPM,
transcript per million; HR, hazard ratio.

Figure 3

Clinical characteristics of GBP5 in
ccRCC. (A) The analysis of GBP5 mRNA expression in tumor and normal
tissues was performed using the Tumor Immune Estimation Resource
2.0. Analysis showed a marked increase in GBP5 expression across a
wide range of malignancies, notably KIRC and KIRP. The association
of GBP5 mRNA expression with different clinical variables in
patients with KIRC, such as (B) T stage, (C) N stage, (D) M stage
and (E) pathological stage. (F) Univariate cox regression analysis
indicates that elevated GBP5 expression is linked to unfavorable
clinical characteristics in patients with ccRCC. (G) Receiver
operating characteristic curve analysis suggests that GBP5 has high
sensitivity and specificity in the diagnosis of ccRCC, with an AUC
of 0.934. *P<0.05, **P<0.01 and ***P<0.001. GBP5,
guanylate binding protein 5; ccRCC, clear cell renal cell
carcinoma; KIRC, kidney renal clear cell carcinoma; KIRP, kidney
renal papillary cell carcinoma; T, tumor; N, lymph node; M,
metastasis; AUC, area under curve; TPM, transcript per million;
TPR, true-positive rate; FDR, false discovery rate.

Figure 4

Analysis of functional enrichment of
GBP5-related DEGs in KIRC. (A) Volcano plot of DEGs related to GBP5
generated using the Cancer Genome Atlas-KIRC dataset
(log2 fold change >2; P<0.05). (B-D) Gene Ontology
enrichment analysis of GBP5-related DEGs. (B) Biological process
analysis. (C) Cellular component analysis. (D) Molecular function
analysis. (E) GeneMANIA protein-protein interaction network for
GBP5 illustrates genes as nodes, with the size of each node
representing the strength of interaction. The most markedly
enriched pathways between the (F) high GBP5 expression and (G) low
GBP5 expression groups determined using gene set enrichment
analysis. GBP5, guanylate-binding protein 5; DEGs, differentially
expressed genes; KIRC, kidney renal clear cell carcinoma; P.adj,
adjusted P-value.

Figure 5

Relationship between GBP5 expression
and immune cell infiltration in KIRC. (A) Analysis of the Tumor
Immune Estimation Resource database revealed a positive correlation
between GBP5 expression and infiltration levels of B cells,
CD8+ T cells, CD4+ T cells, macrophages,
neutrophils and DCs in KIRC. (B and C) The Tumor Immune Single-cell
Hub database was employed to assess the expression of GBP5 in
various tumor microenvironment-associated cellular subpopulations.
In both KIRC GSE111360 and GSE121636 datasets, GBP5 was found to be
highly expressed in CD4+ T cells, proliferating T cells,
regulatory T cells, CD8+ T cells and NK cells. (D) The
Tracking Tumor Immunophenotype database was employed to analyze the
immunological features of GBP5. Results indicate a strong positive
correlation between GBP5 and various immune cells, including
macrophages, NK cells and CD8+ T cells. (E) Western
blotting was used to measure GBP5 and CD163 protein levels in the
samples of patients with ccRCC. The levels of GBP5 and CD163 are
significantly higher in ccRCC tissues compared with adjacent normal
tissues and the relative expression levels of GBP5 and CD163
exhibit a positive correlation. ***P<0.001. GBP5,
guanylate-binding protein 5; KIRC, kidney renal clear cell
carcinoma; ccRCC, clear cell renal cell carcinoma; NK, natural
killer; DC, dendritic cells; TPM, transcript per million, N,
normal; T, tumor.

Figure 6

GBP5 levels in renal cell carcinoma
cells. (A) The detection of GBP5 expression in HK-2, ACHN, 786-O
and Caki-1 cells using western blotting. Results demonstrate that
GBP5 protein levels are higher in renal cancer cell lines compared
with HK-2 cells. (B and C) The efficiency of knockdown and
overexpression of GBP5 in Caki-1 and 786-O cells was determined
using western blotting. Results are presented as mean ± standard
deviation from three distinct experiments. *P<0.05, **P<0.01
and ****P<0.0001. GBP5, guanylate-binding protein 5; OE,
overexpression; sh, short hairpin; NC, negative control; ns, not
significant.

Figure 7

GBP5 enhances clear cell renal cell
carcinoma cell migration and invasion. (A and B) The capability of
cell migration was assessed through wound healing assay (scale bar,
100 µm). Results show that the suppression of GBP5 expression
significantly decreased the cell migratory capacity and GBP5
overexpression enhanced the wound closure rate. Transwell assays
demonstrate that (C) knockdown of GBP5 inhibits cell migration and
invasion, whereas (D) GBP5 overexpression significantly promotes
migration and invasion (scale bar, 100 µm). Results are presented
as mean ± standard deviation from three distinct experiments
*P<0.05, **P<0.01 and ***P<0.001. GBP5, guanylate-binding
protein 5; OE, overexpression; sh, short hairpin; NC, negative
control.
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Copy and paste a formatted citation
Spandidos Publications style
Cheng X, Ye S, Xu X, Yang S, Zhao W, Lu Z and Lai C: GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma. Oncol Lett 30: 571, 2025.
APA
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., & Lai, C. (2025). GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma. Oncology Letters, 30, 571. https://doi.org/10.3892/ol.2025.15317
MLA
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., Lai, C."GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma". Oncology Letters 30.6 (2025): 571.
Chicago
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., Lai, C."GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma". Oncology Letters 30, no. 6 (2025): 571. https://doi.org/10.3892/ol.2025.15317
Copy and paste a formatted citation
x
Spandidos Publications style
Cheng X, Ye S, Xu X, Yang S, Zhao W, Lu Z and Lai C: GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma. Oncol Lett 30: 571, 2025.
APA
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., & Lai, C. (2025). GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma. Oncology Letters, 30, 571. https://doi.org/10.3892/ol.2025.15317
MLA
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., Lai, C."GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma". Oncology Letters 30.6 (2025): 571.
Chicago
Cheng, X., Ye, S., Xu, X., Yang, S., Zhao, W., Lu, Z., Lai, C."GBP5 as a prognostic biomarker associated with immune cell infiltration, migration and invasion in clear cell renal cell carcinoma". Oncology Letters 30, no. 6 (2025): 571. https://doi.org/10.3892/ol.2025.15317
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