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Article Open Access

CIAO1 as a crucial signature gene of cuproptosis in gastric cancer

  • Authors:
    • Jiaying Qu
    • Chunhui Yang
    • Shunchen Zhou
    • Bosen Zhao
    • Qiangsong Tong
    • Liduan Zheng
  • View Affiliations / Copyright

    Affiliations: Department of Pathology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China, Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
    Copyright: © Qu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 440
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    Published online on: July 14, 2025
       https://doi.org/10.3892/ol.2025.15186
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Abstract

Gastric cancer is a global health challenge, necessitating the identification of novel biomarkers and therapeutic targets. The present study aimed to assess the role of cuproptosisrelated genes (CRGs) in gastric cancer, with the goal of establishing a predictive model consisting of key regulators with prognostic significance, thereby enabling the identification of key genes. Data from The Cancer Genome Atlas and Gene Expression Omnibus databases were used to analyze CRGs in stomach adenocarcinoma (STAD). Least absolute shrinkage and selection operator regression analysis was applied to create a risk model, and its predictive accuracy was confirmed for several clinical subgroups. Moreover, the prognostic value of essential regulators was evaluated through multiple analyses. A risk model with 15 CRGs was used to effectively predict STAD prognosis, demonstrating areas under the receiver operating characteristic curve values of 0.822, 0.811 and 0.922 for 1‑, 3‑ and 5‑year overall survival rates, respectively. The risk scores were associated with survival and tumor site. Among the CRGs, the gene for cytosolic iron‑sulfur assembly component 1 (CIAO1) was revealed to be critical and associated with histological type, age and treatment outcome. Moreover, single‑cell analysis demonstrated that CIAO1 is highly expressed in numerous types of cancer cells, and a high expression of CIAO1 was associated with upregulated transcription levels of immune checkpoints, increased tumor mutation load and decreased immune scores, highlighting its complex role in the tumor microenvironment. CIAO1 knockdown experiments were performed, and eliminating CIAO1 was associated with a reduction in the levels of iron‑sulfur proteins and an increase in heat shock protein 70 expression, thereby inducing copper‑dependent cell death. Furthermore, treatment with the drugs dasatinib and AT‑9283 were associated with an inhibition of CIAO1 expression in gastric cancer cells, and decreased rates of tumor spread and invasion. Taken together, the findings of the present study suggest that CIAO1 is a promising biomarker both for assessing prognosis and evaluating the tumor immune microenvironment of gastric cancer.
View Figures

Figure 1

Schematic workflow of the development
and validation of the risk model using TCGA, highlighting key genes
and their interaction mechanisms through gene expression analysis,
survival curve plotting and network diagram construction. Potential
drug targets were identified, and their efficacy was assessed using
in vitro experiments. TCGA, The Cancer Genome Atlas; LASSO,
Least absolute shrinkage and selection operator; ROC, receiver
operating characteristic; PPI, protein-protein interaction; IHC,
immunohistochemistry; RT-qPCR, reverse transcription-quantitative
PCR.

Figure 2

Development of a risk model based on
CRGs in TCGA dataset. (A) A volcano plot was constructed to
identify genes exhibiting differential expression between tumor and
normal tissues (|Log FoldChange|>0.585; P<0.05). (B)
Venn diagram showing 41 DEGs from TGCA-STAD. (C) Least absolute
shrinkage and selection operator coefficient profile. (D) Optimal λ
selection via cross-validation. (E) Survival differences between
high- and low-risk groups were analyzed in the training cohort. (F)
AUC values for 1-, 3- and 5-year survival rates. Receiver operator
characteristic curves were constructed for the different clinical
characteristics, and a risk model was developed. (G) Nomogram was
constructed to predict overall survival rates at 1, 3 and 5 years
for patients with gastric cancer. ***P<0.001. TCGA, The Cancer
Genome Atlas; DEGs, differentially expressed genes; STAD, stomach
adenocarcinoma; CRGs, cuproptosis-related genes; AUC, areas under
the receiver operating characteristic curves.

Figure 3

Clinical characteristics of high- and
low-risk groups of the cuproptosis gene-associated model. The
associations between the risk score derived from the
cuproptosisassociated gene model and several clinical
characteristics of patients with gastric cancer were determined.
The risk score was used to stratify patients into high- and
low-risk groups, and its association with key clinical parameters
was analyzed. (A) OS status; (B) age; (C) sex; (D) grade; (E) T
stage; (F) N stage; (G) M stage; and (H) primary site of patients
with gastric cancer. *P<0.05; **P<0.01. OS, overall survival;
T, tumor; N, lymph node; M, metastasis.

Figure 4

Elevated CIAO1 expression is
crucial for assembly of iron-sulfur clusters in stomach
adenocarcinoma. (A) Heatmap illustrating the correlation among 15
DE-CRGs. (B) Protein-protein interaction network plot of 15
DE-CRGs. (C) Box plot illustrating the z-score of the 15 DE-CRGs.
(D) Mechanism of the iron-sulfur cluster assembly process. (E)
Survival analysis of patients with gastric cancer with high or low
CIAO1 levels, determined using the Kaplan-Meier Plotter
database. (F) CIAO1 gene expression in gastric cancer
tissues compared with adjacent normal tissues. *P<0.05,
**P<0.01, ***P<0.001, ****P<0.0001. (G)
Immunohistochemical staining of CIAO1 in gastric cancer tissue.
Magnification, ×40 times (left) and 400 times (right)
respectively). CIAO1, cytosolic ironsulfur assembly component 1;
DE-CRGs, differentially expressed cuproptosis-related genes; HR,
hazard ratio; N, lymph node; T, tumor; TPM, transcripts per
million.

Figure 5

CIAO1 expression is associated
with clinicopathological features of stomach adenocarcinoma. The
association between the CIAO1 expression levels and several
clinicopathological characteristics of patients with gastric cancer
were determined, providing insights into the potential role of
CIAO1 in tumor progression and patient outcomes. (A)
Treatment outcome; (B) histological type; (C) tumor site; (D) sex;
(E) age; and (F) T stage of patients with gastric cancer.
**P<0.01; ***P<0.001. CIAO1, cytosolic iron-sulfur
assembly component 1; TPM, transcripts per million; T, tumor.

Figure 6

Impact of CIAO1 expression on
the tumor microenvironment and immune-associated markers in stomach
adenocarcinoma. (A) Expression and distribution of CIAO1 across
cell types in gastric cancer. (B) t-SNE plots were used to
visualize the distribution of CIAO1 in cell subtypes in the three
patients with gastric cancer. Expression levels of (C) immune
checkpoints, (D) tumor mutation burden and (E) ESTIMATE scores,
comparing the high- and low-CIAO1 expression groups.
*P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
CIAO1, cytosolic iron-sulfur assembly component 1; tSNE,
t-distributed stochastic neighbor embedding; NK, natural killer;
RBC, red blood cell; pDC, plasmacytoid dendritic cell; TPM,
transcripts per million; PD1, prephenate dehydratase 1;
PDL2, programmed cell death 1 ligand 2; CTLA4,
cytotoxic T-lymphocyte associated protein 4; LAG3,
lymphocyte activating 3; HAVCR2, hepatitis A virus cellular
receptor 2; CEACAM1, carcinoembryonic antigen-related cell
adhesion molecule 1; SIRPA, signal regulatory protein α; ns,
not significant.

Figure 7

Enrichment analysis of DEGs and
CIAO1-interacting proteins in stomach adenocarcinoma. (A) Volcano
plot, highlighting the DEGs between the high- and low-CIAO1
expression groups [Log(FoldChange)<1; P<0.05]. (B)
Gene Ontology functional enrichment analysis for DEGs. (C) Geneset
enrichment analysis. (D) Protein-protein interaction network plot
of CIAO1-interacting proteins. (E) Kyoto Encyclopedia of Genes and
Genomes functional enrichment analysis for DEGs. The
three-dimensional visualization of the structure of CIAO1 and its
interacting proteins: (F) CIAO2A, (G) CIAO2B and (H) CIAO3. DEGs,
differentially expressed genes; CIAO, cytosolic iron-sulfur
assembly component; BP, Biological Process; CC, Cellular
Compartment; MF, Molecular Function.

Figure 8

Identification of transcriptional
inhibitors of CIAO1 expression. Drug sensitivity assessments
for (A) dasatinib, (B) AT-9283, (C) vorinostat, (D) idebenone, (E)
ZM-336372, (F) methylprednisolone, (G) econazole nitrate, (H)
masoprocol, (I) fludarabine, (J) cladribine, (K) nitazoxanide and
(L) nelarabine on CIAO1 transcript levels. CIAO1,
cytosolic ironsulfur assembly component 1.

Figure 9

Pharmacological inhibition of CIAO1
induces cuproptosis and inhibits the aggressiveness of cancer
cells. (A) Western blot analysis revealed the expression of CIAO1
in GES 1, AGS, SNU 1, MKN 45, HGC 27, HEK293 and HeLa cell lines.
(B) RT-qPCR assays (normalized to βactin) demonstrated the
CIAO1 expression levels, (C) western blot analysis revealed
the CIAO1, FDX1, LIAS and HSP70 expression levels and (D)
immunofluorescence assays demonstrated the localization and
fluorescence intensity of CIAO1 expression (scale bar, 10 µm) in
AGS and MKN-45 cells transfected with sh-Scb, sh-CIAO1 #1 or
sh-CIAO1 #2. (E) Western blot analysis revealed the CIAO1,
FDX1, LIAS and HSP70 expression levels in AGS and MKN-45 cells
treated with dasatinib (0.5 nmol/l) or AT-9283 (4 nmol/l). MTT
colorimetric assays demonstrated changes in the viability of AGS
and MKN-45 cells (F) transfected with sh-Scb, sh-CIAO1 #1 or
sh-CIAO1 #2 and (G) treated with dasatinib (0.5 nmol/l) or
AT9283 (4 nmol/l). Representative images (left panels) and
quantification of the data (right panels) of (H) soft agar and (I)
Matrigel invasion assays, demonstrated the anchorage-independent
proliferation and invasion of AGS and MKN-45 cells stably
transfected with sh-Scb, sh-CIAO1 #1 or sh-CIAO1 #2.
Magnification, ×100. ***P<0.001; ****P<0.0001 vs.
sh-Scb. CIAO1, cytosolic iron-sulfur assembly component 1;
RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin;
Scb, scramble; FDX1, ferredoxin 1; LIAS, lipoic acid synthetase;
HSP70, heat shock protein 70; ns, not significant.
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Copy and paste a formatted citation
Spandidos Publications style
Qu J, Yang C, Zhou S, Zhao B, Tong Q and Zheng L: CIAO1 as a crucial signature gene of cuproptosis in gastric cancer. Oncol Lett 30: 440, 2025.
APA
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., & Zheng, L. (2025). CIAO1 as a crucial signature gene of cuproptosis in gastric cancer. Oncology Letters, 30, 440. https://doi.org/10.3892/ol.2025.15186
MLA
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., Zheng, L."CIAO1 as a crucial signature gene of cuproptosis in gastric cancer". Oncology Letters 30.3 (2025): 440.
Chicago
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., Zheng, L."CIAO1 as a crucial signature gene of cuproptosis in gastric cancer". Oncology Letters 30, no. 3 (2025): 440. https://doi.org/10.3892/ol.2025.15186
Copy and paste a formatted citation
x
Spandidos Publications style
Qu J, Yang C, Zhou S, Zhao B, Tong Q and Zheng L: CIAO1 as a crucial signature gene of cuproptosis in gastric cancer. Oncol Lett 30: 440, 2025.
APA
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., & Zheng, L. (2025). CIAO1 as a crucial signature gene of cuproptosis in gastric cancer. Oncology Letters, 30, 440. https://doi.org/10.3892/ol.2025.15186
MLA
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., Zheng, L."CIAO1 as a crucial signature gene of cuproptosis in gastric cancer". Oncology Letters 30.3 (2025): 440.
Chicago
Qu, J., Yang, C., Zhou, S., Zhao, B., Tong, Q., Zheng, L."CIAO1 as a crucial signature gene of cuproptosis in gastric cancer". Oncology Letters 30, no. 3 (2025): 440. https://doi.org/10.3892/ol.2025.15186
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