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Article Open Access

FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages

  • Authors:
    • Liangliang Bai
    • Jing Xiong
    • Sihai Chen
    • Jiahao Hu
    • Meixia Zhang
    • Bimin Li
    • Jing Hu
    • Mingyan He
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, Jiangxi Provincial Key Laboratory of Digestive Diseases, Jiangxi Clinical Research Center for Gastroenterology, Digestive Disease Hospital, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of General Practice, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Cardiology, Jiangxi Provincial People's Hospital, The First Affiliated Hospital of Nanchang Medical College, Nanchang, Jiangxi 330200, P.R. China
    Copyright: © Bai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 36
    |
    Published online on: December 2, 2025
       https://doi.org/10.3892/ijmm.2025.5707
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Abstract

Macrophages play a key role in hepatocellular carcinoma (HCC) progression, but the mechanisms underlying this involvement remain unclear. In the present study, mice with HCC were used for in vivo experiments, and 97H and THP‑1 cells were used for in vitro experiments. Metabolomic analysis was performed to detect changes of metabolites in the supernatant of 97H cells. Flow cytometry and immunohistochemical staining were performed to assess macrophage polarization. Western blotting was performed to examine the levels of phosphorylated (p‑) PI3K, p‑AKT and NRF2. Reverse transcription‑quantitative polymerase chain reaction was performed to examine FBXO22, IMPA1 and PTEN mRNA expression levels. FBXO22 significantly promoted the release of myo‑inositol in the cell supernatant of 97H cells, markedly decreased the number of CD86‑positive cells (M1 macrophages), and increased the number of CD206‑positive cells (M2 macrophages) in both THP‑1 cells and mouse HCC tumor tissues. The promoting effect of myo‑inositol on M2 macrophages was reversed by transfection with small interfering (si)‑SLC5A3 in vitro. In addition, FBXO22 overexpression reduced PTEN protein levels and then elevated NRF2 protein levels upregulating IMPA1 and inducing myo‑inositol release in 97H cells. Co‑culturing of 97H and THP‑1 cells revealed that the stimulatory effect of 97H cells transfected with an overexpression (oe)‑FBXO22 construct on M2 macrophages was reversed by co‑transfection with the si‑IMPA1. Co‑immunoprecipitation revealed a promoting effect of FBXO22 on PTEN ubiquitination via direct interaction in 97H cells. Furthermore, luciferase activity and chromatin immunoprecipitation assays indicated direct transcriptional regulation of IMPA1 expression by NRF2 in 97H cells. The in vivo experiments further revealed that transfection with the si‑IMPA1 reversed the promoting effect of oe‑FBXO22 on tumor growth and M2 polarization by reducing myo‑inositol levels in tumor tissues. In conclusion, FBXO22 degrades PTEN by inducing its ubiquitination to elevate NRF2 protein levels. As a result, IMPA1 expression is increased, which causes myo‑inositol release by HCC cells and further induces M2‑type macrophages via SLC5A3 to promote HCC tumor growth. The present study identified a novel molecular mechanism by which FBXO22 promotes HCC progression.
View Figures

Figure 1

Detection of differentially expressed
metabolites in 97H cells overexpressing FBXO22. 97H cells
were transfected with oe-FBXO22, and cell supernatants were
collected for non-target metabolomic analysis. Hierarchical
clustering results of the top 50 differential metabolites (VIP
>1) are illustrated. **P<0.01, ***P<0.001. Oe-,
overexpression.

Figure 2

FBXO22-induced myo-inositol release
promotes M2 polarization of THP-1 cells. 97H cells were treated
with myo-inositol. (A) A Cell Counting Kit-8 assay was performed to
assess cell viability. (B and C) Flow cytometry was performed to
detect CD86- and CD206-positive THP-1 cells. (D and E) Western
blotting was performed to examine p-PI3K and p-AKT levels in THP-1
cells. *P<0.05 and ***P<0.001. p-,
phosphorylated.

Figure 3

Myo-inositol induces M2 polarization
of THP-1 cells via SLC5A3. THP-1 cells were transfected with
si-SLC5A3. (A) Reverse transcription-quantitative PCR and
(B) western blotting were performed to detect SLC5A3 expression.
THP-1 cells were transfected with si-SLC5A3, followed by
treatment with 10 μM myo-inositol. (C and D) Flow cytometry
was performed to detect CD86-positive and CD206-positive THP-1
cells. (E and F) Western blotting was performed to examine p-PI3K
and p-AKT levels in THP-1 cells. *P<0.05,
**P<0.01 and ***P<0.001. p-,
phosphorylated.

Figure 4

FBXO22 induces PTEN ubiquitination
and subsequent degradation in THP-1 cells. 97H cells were
transfected with oe-FBXO22. Reverse
transcription-quantitative PCR and western blotting were performed
to detect mRNA expression levels and protein levels of (A, C and D)
FBXO22, (B, C and E) IMPA1 and (G and H) PTEN.
(F) A myo-inositol detection kit was used to detect myo-inositol in
cell supernatants. (I and J) 97H cells were transfected with
oe-FBXO22, followed by the treatment with CHX (200
μg/ml) for different periods (2, 4 and 8 h). Western
blotting was performed to detect PTEN protein levels. (K) 97H cells
were transfected with oe-FBXO22 followed by the treatment
with MG132 (10 μM) for 12 h. Western blotting was performed
to detect PTEN protein levels. (L) 97H cells were incubated with an
anti-PTEN antibody, and western blotting was performed to detect
FBXO22 protein levels. (M) 97H cells were incubated with an
anti-PTEN antibody, followed by the transfection with
oe-FBXO22. Western blotting was performed to detect PTEN
ubiquitination levels. *P<0.05,
**P<0.01 and ***P<0.001. CHX,
cycloheximide; oe-, overexpression; NC, negative control.

Figure 5

FBXO22-induced myo-inositol release
promotes M2 polarization via the regulation of the NRF2/IMPA1 axis
in THP-1 cells. (A) 97H cells were incubated with an anti-NRF2
antibody, and western blotting was performed to detect PTEN protein
levels. (B) 97H cells were transfected with oe-FBXO22, and
western blotting was performed to detect NRF2 protein level. (C)
293T cells were transfected with oe-NRF2, and luciferase
assay was performed to determine the interaction between NRF2 and
IMPA1. (D) Chromatin immunoprecipitation assay was used to detect
the transcriptional regulation of IMPA1 by NRF2 in 97H cells
transfected with oe-FBXO22, IgG was used as negative
control. 97H cells were co-transfected with oe-FBXO22 and
si-IMPA1. (E and F) Reverse transcription-quantitative PCR
and western blotting were performed to detect IMPA1
expression. (G) A myo-inositol detect kit was used to detect
myo-inositol in cell supernatants. (H and I) Flow cytometry was
performed to detect CD86-positive and CD206-positive THP-1 cells.
**P<0.01 and ***P<0.001. oe-,
overexpression; si-, small interfering; NC, negative control.

Figure 6

Myo-inositol promotes HCC tumor
growth and induces M2 polarization in vivo. BALB/c nude mice
received a subcutaneous injection of 97H cells to induce HCC. The
mice were randomly divided into two groups (n=4 each): PBS group
and myo-inositol group. (A and B) Tumor growth was monitored every
3 days for ~3 weeks. (C and D) Western blotting was performed to
detect p-PI3K and p-AKT levels in tumor tissues. (E and F) IHC
staining was performed to detect CD86-positive and CD206-positive
cells in tumor tissues. *P<0.05 and
**P<0.01. HCC, hepatocellular carcinoma; p-,
phosphorylated.

Figure 7

Transfection with si-IMPA1
reverses the stimulatory effect of FBXO22 on tumor growth and M2
polarization in vivo. BALB/c nude mice were subcutaneously
injected with 97H cells to induce hepatocellular carcinoma. The
model mice were randomly divided into four groups (n=4 each):
97H-NC + si-NC, 97H-FBXO22 + si-NC, 97H-NC + si-IMPA1, and
97H-FBXO22 + si-IMPA1. (A and B) Tumor growth was monitored every 3
days for ~3 weeks. (C) A myo-inositol detection kit was used to
detect myo-inositol in tumor tissues. (D) Reverse
transcription-quantitative PCR was performed to detect IMPA1 mRNA
expression in tumor tissues. (E) Western blotting was performed to
detect IMPA1 protein level in tumor tissues. (F and G) IHC staining
was performed to detect CD86-positive and CD206-positive cells in
tumor tissues. **P<0.01 and ***P<0.001.
si-, small interfering; NC, negative control.
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Copy and paste a formatted citation
Spandidos Publications style
Bai L, Xiong J, Chen S, Hu J, Zhang M, Li B, Hu J and He M: FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages. Int J Mol Med 57: 36, 2026.
APA
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B. ... He, M. (2026). FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages. International Journal of Molecular Medicine, 57, 36. https://doi.org/10.3892/ijmm.2025.5707
MLA
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B., Hu, J., He, M."FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages". International Journal of Molecular Medicine 57.2 (2026): 36.
Chicago
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B., Hu, J., He, M."FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages". International Journal of Molecular Medicine 57, no. 2 (2026): 36. https://doi.org/10.3892/ijmm.2025.5707
Copy and paste a formatted citation
x
Spandidos Publications style
Bai L, Xiong J, Chen S, Hu J, Zhang M, Li B, Hu J and He M: FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages. Int J Mol Med 57: 36, 2026.
APA
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B. ... He, M. (2026). FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages. International Journal of Molecular Medicine, 57, 36. https://doi.org/10.3892/ijmm.2025.5707
MLA
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B., Hu, J., He, M."FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages". International Journal of Molecular Medicine 57.2 (2026): 36.
Chicago
Bai, L., Xiong, J., Chen, S., Hu, J., Zhang, M., Li, B., Hu, J., He, M."FBXO22 promotes hepatocellular carcinoma progression via paracrine myo‑inositol‑induced M2‑type polarization of macrophages". International Journal of Molecular Medicine 57, no. 2 (2026): 36. https://doi.org/10.3892/ijmm.2025.5707
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