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ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis

  • Authors:
    • Liyun Yang
    • Junzhi Liu
    • Yuhuan Gao
    • Shuixian Huang
    • Geping Wu
  • View Affiliations / Copyright

    Affiliations: Department of Otolaryngology, Gongli Hospital of Shanghai Pudong New Area, Shanghai 200135, P.R. China, Department of Clinical Medicine, Medical College of Yangzhou University, Yangzhou, Jiangsu 225009, P.R. China, Department of Otolaryngology, Zhangjiagang Hospital Affiliated to Soochow University, Suzhou, Jiangsu 215600, P.R. China
    Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 137
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    Published online on: June 12, 2025
       https://doi.org/10.3892/br.2025.2015
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Abstract

Thyroid cancer (THCA) is a prevalent malignancy of the head and neck region, yet the mechanisms underlying its tumorigenesis and metastasis remain poorly understood. Given that Rho GTPase activating protein 36 (ARHGAP36) has been implicated in various cellular processes related to cancer progression, including cell migration and invasion, it represents a promising candidate for further investigation in THCA. To investigate the gene expression differences in ARHGAP36 between tumor and normal tissues, the GEPIA and UALCAN databases were utilized. Various factors were also evaluated, including sample types, cancer stages, demographics, histological characteristics and nodal status. The LinkedOmics database was used to constructed a co‑expression network for ARHGAP36 in THCA. Gene Ontology (GO) process enrichment analysis for ARHGAP36‑associated genes was conducted via Metascape. The TIMER and TISCH databases were employed to explore the relationships between ARHGAP36 and immune markers as well as cell clusters. Functional in vitro assays were performed to assess cellular behaviors such as proliferation, migration and apoptosis. The results indicated that ARHGAP36 expression was significantly elevated in THCA tissues compared with normal tissues. Co‑expression analysis revealed significant links between ARHGAP36 and key genes, including IGSF1, DPYSL3, ZCCHC12, CD97, LOXL4, CST2 and WSCD. The enriched GO processes involved T‑cell immunity, particularly highlighting the association between ARHGAP36 and CD4+ T cell infiltration. Notably, the downregulation of ARHGAP36 reduced tumor cell proliferation, migration and invasion while enhancing apoptosis. In conclusion, the findings of the present study indicate that ARHGAP36 plays a crucial role in facilitating immune evasion and promoting THCA progression. This underscores its potential as a diagnostic marker and therapeutic target in THCA.
View Figures

Figure 1

ARHGAP36 expression is higher in THCA
samples than in normal tissues. GEPIA database analyses of ARHGAP36
mRNA expression in (A) TCGA and (B) TCGA-THCA tumor tissues
compared with adjacent normal tissues. (C) ARHGAP36 expression in
TCGA tumor and normal tissues. (D) Differential ARHGAP36 mRNA
expression in the THCA dataset (TCGA). *P<0.05,
**P<0.01. ARHGAP36, Rho GTPase activating protein 36;
THCA, thyroid cancer; TCGA, The Cancer Genome Atlas; TPM,
transcripts per million.

Figure 2

Higher ARHGAP36 mRNA expression is
associated with TNM stage in patients with THCA. (A) ARHGAP36
expression in tumor tissues is significantly higher than in normal
tissues. (B) ARHGAP36 expression increases across tumor stages 1-4.
Elevated ARHGAP36 expression by (C) ethnicity, (D) sex, (E) age and
(F) THCA subtype. (G) Higher ARHGAP36 expression in patients with
lymph node metastasis. *P<0.05,
**P<0.01. The UALCAN data, which was used for all
analyses, revealed a comprehensive association between ARHGAP36
expression and various clinical parameters in THCA. ARHGAP36, Rho
GTPase activating protein 36; THCA, thyroid cancer; TCGA, The
Cancer Genome Atlas.

Figure 3

ARHGAP36 downregulation reduces
proliferation and induces apoptosis in THCA cells. (A) Western blot
assays show the effectiveness of the siRNA transfection and confirm
the significant decrease in ARHGAP36 protein levels. (B) Protein
ratio in cells. (C) Colony formation assays show reduced
proliferation in BCPAP/si-ARHGAP36 cells. (D) Colony numbers for
BCPAP/si-ARHGAP36 cells. (E) Colony formation assays show reduced
proliferation in BHT101/si-ARHGAP36 cells. (F) Colony numbers for
BHT101/si-ARHGAP36 cells. (G) Apoptosis assays show ARHGAP36
downregulation induces the apoptosis of BCPAP. (H) Apoptosis rates
in BCPAP cells. (I) Apoptosis assays show ARHGAP36 downregulation
the induces apoptosis of BHT101 cells. (J) Apoptosis rates in
BHT101 cells. **P<0.01. ARHGAP36, Rho GTPase
activating protein 36; THCA, thyroid cancer; NC, negative control;
siRNA, small interfering RNA.

Figure 4

ARHGAP36 downregulation reduces the
migration and invasion of thyroid cancer cells. Wound healing
assays reveal reduced movement in ARHGAP36 knockdown (A and C)
BCPAP/si-ARHGAP36 and (B and D) BHT101/si-ARHGAP36 cells. Transwell
migration assays show decreased migration in (E and G)
BCPAP/si-ARHGAP36 and (F and H) BHT101/si-ARHGAP36 cells. Transwell
invasion assays indicate reduced invasion in (E and G)
BCPAP/si-ARHGAP36 and (F and H) BHT101/si-ARHGAP36 cells.
*P<0.05, **P<0.01. ARHGAP36, Rho GTPase
activating protein 36; si, small interfering (RNA); NC, negative
control.

Figure 5

ARHGAP36 gene co-expression network in
thyroid cancer. (A) Volcano plot of genes co-expressed with
ARHGAP36 in TCGA. Heatmaps of the top 50 (B) positively and (C)
negatively correlated genes. Positive correlations between ARHGAP36
and (D) IGSF1, (E) DPYSL3 and (F) ZCCHC12. Negative correlations
with (G) SETD3, (H) COX15 and (I) RNF157. ARHGAP36, Rho GTPase
activating protein 36; IGSF1, immunoglobulin superfamily member 1;
DPYSL3, dihydropyrimidinase like 3; ZCCHC12, zinc finger CCHC-type
containing 12; TPM, transcripts per million. SETD3, SET
domain-containing protein 3; RNF157, ring finger protein 157.

Figure 6

T cell-mediated immunity is
significantly enriched among genes related to ARHGAP36. (A) Gene
Ontology biological processes enriched in ARHGAP36-associated
genes. (B,C) Protein-protein interaction networks with functional
clusters represented by color. ARHGAP36, Rho GTPase activating
protein 36.

Figure 7

ARHGAP36 weakly correlates with immune
CD4+ T cell infiltration in thyroid cancer. TIMER
analysis of ARHGAP36 correlations with (A) purity, (B) B cells, (C)
CD8+ T cells, (D) CD4+ T cells, (E)
macrophages, (F) neutrophils and (G) dendritic cells. ARHGAP36, Rho
GTPase activating protein 36; TPM, transcripts per million.

Figure 8

ARHGAP36 is associated with immune
escape. (A,B) Single-cell expression matrices from the TISCH
database show ARHGAP36 abundance in immune cell clusters. Positive
associations between ARHGAP36 and immune checkpoints, (C) PDCD-1,
(D) LAG3, (E) CD274 and (F) CTLA4. ARHGAP36, Rho GTPase activating
protein 36; TPM, transcripts per million; LAG3,
lymphocyte-activation gene 3; CTLA4, cytotoxic
T-lymphocyte-associated protein 4.
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Copy and paste a formatted citation
Spandidos Publications style
Yang L, Liu J, Gao Y, Huang S and Wu G: ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis. Biomed Rep 23: 137, 2025.
APA
Yang, L., Liu, J., Gao, Y., Huang, S., & Wu, G. (2025). ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis. Biomedical Reports, 23, 137. https://doi.org/10.3892/br.2025.2015
MLA
Yang, L., Liu, J., Gao, Y., Huang, S., Wu, G."ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis". Biomedical Reports 23.2 (2025): 137.
Chicago
Yang, L., Liu, J., Gao, Y., Huang, S., Wu, G."ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis". Biomedical Reports 23, no. 2 (2025): 137. https://doi.org/10.3892/br.2025.2015
Copy and paste a formatted citation
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Spandidos Publications style
Yang L, Liu J, Gao Y, Huang S and Wu G: ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis. Biomed Rep 23: 137, 2025.
APA
Yang, L., Liu, J., Gao, Y., Huang, S., & Wu, G. (2025). ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis. Biomedical Reports, 23, 137. https://doi.org/10.3892/br.2025.2015
MLA
Yang, L., Liu, J., Gao, Y., Huang, S., Wu, G."ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis". Biomedical Reports 23.2 (2025): 137.
Chicago
Yang, L., Liu, J., Gao, Y., Huang, S., Wu, G."ARHGAP36 serves as a diagnostic and therapeutic marker that mediates immune escape and promotes thyroid cancer metastasis". Biomedical Reports 23, no. 2 (2025): 137. https://doi.org/10.3892/br.2025.2015
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