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Article Open Access

GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway

  • Authors:
    • Guangchuan Mu
    • Kaiyan Li
    • Chaozhen Hu
    • Jintao Cai
    • Huichao Ruan
    • Guanyu Zhu
    • Dang Liu
  • View Affiliations / Copyright

    Affiliations: Department of Gastrointestinal Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
    Copyright: © Mu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 48
    |
    Published online on: January 19, 2026
       https://doi.org/10.3892/or.2026.9053
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Abstract

Gastric cancer is a malignancy with a high incidence and poor prognosis. The identification of novel molecular markers and elucidation of their underlying mechanisms may provide new avenues for improving therapeutic strategies. The present study analyzed the association between GPR176 expression and clinicopathological features using The Cancer Genome Atlas‑Stomach Adenocarcinoma and GSE66254 datasets, and further validated the findings in patients from The First Affiliated Hospital of Guangxi Medical University (Nanning, China). The migratory and invasive abilities of gastric cancer cells were assessed using Transwell and wound‑healing assays. Western blotting was carried out to evaluate the effects of GPR176 on the PI3K/AKT/mTOR signaling pathway. In vivo tumorigenesis assays in nude mice were carried out to confirm the role of GPR176 in tumor progression. Analysis revealed that GPR176 expression was significantly elevated in gastric cancer tissues and associated with unfavorable patient outcomes. Silencing GPR176 markedly suppressed the migration and invasion of gastric cancer cells, accompanied by inhibition of the PI3K/AKT/mTOR and EMT signaling pathways. These inhibitory effects were prevented by the overexpression of PIP5K1A. In line with the in vitro results, experiments with nude mice demonstrated that GPR176 knockdown impeded tumor growth, whereas its overexpression enhanced tumorigenicity. Furthermore, GPR176 suppression significantly attenuated EMT and PI3K/AKT/mTOR signaling in vivo, while GPR176 overexpression led to activation of these pathways. In summary, the present study identifies GPR176 as a novel prognostic biomarker in gastric cancer. Mechanistically, GPR176 promotes EMT and tumor progression, at least in part, through activation of the PI3K/AKT/mTOR signaling pathway.

View Figures

Figure 1

GPR176 expression levels were
associated with clinicopathological characteristics and prognosis
in STAD patients. (A) GPR176 expression levels in common malignant
tumors and adjacent tissues. (B) Comparison of GPR176 expression
levels in STAD tumor tissues and adjacent normal stomach tissues.
(C) Expression levels of GPR176 mRNA in STAD tumors after TNM
staging (I, II, III and IV). (D) Expression levels of GPR176 mRNA
in STAD tumors at pathologic T1, T2, T3 and T4 stages. (E) Tn the
TCGA-STAD dataset, OS of patients with STAD with GPR176 high and
low expression groups using the median value as a cut-off. (F) In
the GSE66254 dataset, OS of patients with STAD with GPR176 high and
low expression groups using the 75th percentile value as a cut-off.
(G) In the GSE66254 dataset, RFS of patients with STAD with GPR176
high and low expression groups using the 75th percentile as a
cut-off. (H) Association of GPR176 expression with pathologic T
staging in the TCGA-STAD dataset. (I) The ROC curve based on GPR176
plotted for distinguishing between gastric cancer and adjacent
tissues. (J) Significant positive correlation between GPR176 and
PIP5K1A expression in the gastric cancer samples in the TCGA-STAD
dataset. ROC, receiver operating characteristic; TCGA, The Cancer
Genome Atlas; STAD, stomach adenocarcinoma; OS, overall survival;
RFS, Recurrence-free survival. *P<0.05, **P<0.01,
***P<0.001. Unpaired Student's t-test was used in Fig. 1A-D, Survival analysis with
Kaplan-Meier method was used in Fig.
1E-G; Chi-squared test was used in Fig. 1F; Receiver Operating Characteristic
(ROC) was used in Fig. 1G; Linear
regression was used in Fig. 1H.

Figure 2

The Nomogram was constructed based on
GPR176 and clinical-pathological features in the TCGA-STAD dataset.
(A) Nomogram; (B) fitted curve evaluating the accuracy of on1-year
survival prediction; (C) fitted curve evaluating the accuracy of
3-year survival prediction; (D) fitted curve evaluating the
accuracy of 5-year survival prediction.

Figure 3

Bioinformatics analysis revealed
potential mechanisms through which GPR176 may exert its functions.
(A) Volcano plot of differentially expressed genes between GPR176
high and low expression groups in the TCGA-STAD dataset. (B) KEGG
signaling pathways enriched based on differentially expressed
genes. (C) GO terms enriched based on differentially expressed
genes. (D) Gene Set Enrichment Analysis results based on
differentially expressed genes. (E) Correlation between GPR176
expression and the expression of target genes of PI3K/AKT/mTOR
signaling pathway, including CDK2, FOXO1, FOXO3 and VEGFA. GO, Gene
Ontology; BP, biological process; CC, cellular component; MF,
molecular function; KEGG, Kyoto Encyclopedia of Genes and Genomes.
***P<0.001.

Figure 4

Genetic regulation of GPR176
influences cell invasion/migration and EMT signaling pathways in
gastric cancer cells. (A) Upregulation and downregulation of GPR176
promote and inhibit migration of gastric cancer cells,
respectively. (B) Upregulation and downregulation of GPR176 promote
and inhibit invasion of gastric cancer cells, respectively. (C)
Effects of upregulation and downregulation of GPR176 on mRNA
expression of EMT pathway genes in HGC-27 and NCI-N87 cells. (D)
Effects of upregulation and downregulation of GPR176 on protein
expression in the EMT pathway in HGC-27 and NCI-N87 cells, with the
corresponding bar chart shown in Fig.
S2A and B. EMT, epithelial-mesenchymal transition; sh-NC, short
hairpin negative control; cad, cadherin. Statistical analysis was
carried out using one-way ANOVA. ***P<0.001, **P<0.01.

Figure 5

Genetic regulation of PIP5K1A
counteracts the effects of GPR176 on invasion, migration. (A)
Combined genetic regulation of GPR176 and PIP5K1A on cell invasion.
(B) Combined genetic regulation of GPR176 and PIP5K1A on cell
migration. sh-NC, short hairpin-negative control. Statistical
analysis was carried out using two-way ANOVA. *P<0.05,
**P<0.01.

Figure 6

Genetic regulation of PIP5K1A
counteracts the effects of GPR176 on EMT pathway and PI3K/AKT/mTOR
pathway. (A) Combined genetic regulation of GPR176 and PIP5K1A on
protein expression and phosphorylation in the EMT pathway and
PI3K/AKT/mTOR pathway in HGC-27 cells, with the corresponding bar
chart shown in Fig. S3A. (B)
Combined genetic regulation of GPR176 and PIP5K1A on protein
expression and phosphorylation in the EMT pathway and PI3K/AKT/mTOR
pathway in HGC-27 cells, with the corresponding bar chart shown in
Fig. S3B. EMT,
epithelial-mesenchymal transition; sh, short hairpin-; Lenti,
Lentivirus; cad, cadherin.

Figure 7

Genetic regulation of PIP5K1A
counteracts the effects of GPR176 on the expression of EMT pathway
genes at the mRNA level. Combined genetic regulation of GPR176 and
PIP5K1A on the expression of (A) GPR176 (B) CHD1 (E-cadherin), (C)
CHD2 (N-cadherin) (D) VIM (Vimentin), (E) SNAI1 (Snail), (F)
PIP5K1A in in HGC-27 cells. Combined genetic regulation of GPR176
and PIP5K1A on the expression of (G) GPR176, (H) CHD1 (E-cadherin),
(I) CHD2 (N-cadherin), (J) VIM (Vimentin), (K) SNAI1 (Snail), (L)
PIP5K1A in NCI-N87 cells. EMT, epithelial-mesenchymal transition;
sh-, short hairpin-; cad, cadherin. Statistical analysis was
performed using two-way ANOVA. **P<0.01, *P<0.05.

Figure 8

A significant difference was observed
in the expression of key molecules in the EMT and PI3K/AKT/mTOR
pathways, as well as in GPR176 and PIP5K1A levels, between
metastatic and non-metastatic tumor tissues. (A) The relative
expression of E-cadherin at the protein level in the specific
group. (B) Relative expression of GPR176 at the protein level in
the specific group. (C) Relative expression of N-cadherin at the
protein level in the specific group. (D) Relative expression of
PIP5K1A at the protein level in the specific group. (E) Relative
expression of p-AKT/AKT at the protein level in the specific group.
(F) Relative expression of p-mTOR/mTOR at the protein level in the
specific group. (G) The representative blots of key molecules of
the EMT signaling pathway and PI3K/AKT/mTOR pathway at the protein
level in the specific group, as well as the expression of GPR176
and PIP5K1A. (H) Relative expression of CDH1 (E-cadherin) at the
mRNA level in the specific group. (I) Relative expression of GPR176
at the mRNA level in the specific group. (J) Relative expression of
PIK5K1A at the mRNA level in the specific group. (K) Relative
expression of CDH2 (N-cadherin) at the mRNA level in the specific
group. (L) Relative expression of CDH1 (E-cadherin) at the mRNA
level in the specific group. (M) Relative expression of GPR176 at
the mRNA level in the specific group. (N) Relative expression of
PIK5K1A at the mRNA level in the specific group. (O) Relative
expression of CDH2 (N-cadherin) at the mRNA level in the specific
group. Para, para-cancer. For experiments in Fig. 8A-J, statistical analysis was
performed using two-way ANOVA. For the experiments presented in
Fig. 8K-R, an unpaired Student's
t-test was used to assess the statistical significance of
differences between the two groups. **P<0.01.

Figure 9

Genetic regulation of GPR176
expression affects mouse subcutaneous graft tumor growth,
PI3K/AKT/mTOR pathway and the EMT pathway. (A) Effects of
upregulation and knockdown of GPR176 expression on tumor growth.
(B) Effects of GPR176 expression regulation on the expression of
PIP5K1A and EMT signaling pathway genes. (C) Effects of GPR176
expression regulation on the EMT signaling pathway and
PI3K/AKT/mTOR pathway. SH-, short hairpin-; NC, negative control;
EMT, epithelial-mesenchymal transition; cad, cadherin. Statistical
analysis was performed using an unpaired Student's t-test.
**P<0.01, ***P<0.001.
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Copy and paste a formatted citation
Spandidos Publications style
Mu G, Li K, Hu C, Cai J, Ruan H, Zhu G and Liu D: <p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>. Oncol Rep 55: 48, 2026.
APA
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., & Liu, D. (2026). <p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>. Oncology Reports, 55, 48. https://doi.org/10.3892/or.2026.9053
MLA
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., Liu, D."<p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>". Oncology Reports 55.3 (2026): 48.
Chicago
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., Liu, D."<p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>". Oncology Reports 55, no. 3 (2026): 48. https://doi.org/10.3892/or.2026.9053
Copy and paste a formatted citation
x
Spandidos Publications style
Mu G, Li K, Hu C, Cai J, Ruan H, Zhu G and Liu D: <p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>. Oncol Rep 55: 48, 2026.
APA
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., & Liu, D. (2026). <p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>. Oncology Reports, 55, 48. https://doi.org/10.3892/or.2026.9053
MLA
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., Liu, D."<p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>". Oncology Reports 55.3 (2026): 48.
Chicago
Mu, G., Li, K., Hu, C., Cai, J., Ruan, H., Zhu, G., Liu, D."<p>GPR176 enhances the epithelial‑mesenchymal transition in gastric cancer cells by activating the PI3K/AKT/mTOR signaling pathway</p>". Oncology Reports 55, no. 3 (2026): 48. https://doi.org/10.3892/or.2026.9053
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