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HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation

  • Authors:
    • Chenning Zhang
    • Yu Jiang
    • Jiahui Liu
    • Wenyu Zhang
    • Jie Qi
    • Qing Wen
    • Hengli Zhao
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    Affiliations: Department of Clinical Research Center, Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250013, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 47
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    Published online on: January 19, 2026
       https://doi.org/10.3892/or.2026.9052
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Abstract

Hepatocellular carcinoma (HCC) represents the most common form of primary liver cancer and is characterized by a significant rate of recurrence. However, there is still a lack of effective therapeutic methods. Accumulating evidence has highlighted the importance of homeobox containing 1 (HMBOX1) in tumorigenesis. However, the relationship between HMBOX1 expression and HCC remains unclear. In the present study, through the analysis of public databases and staining analysis of tissue microarrays, it was found that compared with normal tissues, HMBOX1 was significantly downregulated in tumor tissues. Furthermore, through analyses such as Cell Counting Kit‑8 assay, wound healing assay and colony formation, it was found that overexpression of HMBOX1 could inhibit cell proliferation and migration, while silencing of HMBOX1 promoted tumor biological characteristics in HCC cell lines. The molecular biological mechanism was explored by using proteomics combined with bioinformatics analysis and western blotting. Mechanistically, AKT1 was identified as a downstream effector of HMBOX1, and protein tyrosine phosphatase non‑receptor type 1 (PTPN1) signaling might mediate the regulation of AKT1 by HMBOX1. In vivo tumor‑bearing experiments also verified the function of the HMBOX1/PTPN1/AKT1 pathway in HCC development. Taken together, the present findings revealed a new HMBOX1/PTPN1/AKT1 axis that inhibits tumor progression and provides new candidate therapy targets for HCC.

View Figures

Figure 1

HMBOX1 expression is downregulated in
tumor tissues and correlates with poor prognosis in patients with
HCC. (A) The expression of HMBOX1 in HCC tumor tissues was analyzed
using the GEPIA2 database. (B) The relationship between HMBOX1
expression and survival was analyzed via Kaplan-Meier plotter
database. (C) IHC analysis of HMBOX1 expression in HCC tumor
tissues and adjacent non-tumor tissues from 48 patients with HCC
using TMA. **P<0.01. HMBOX1, homeobox containing 1; HCC,
hepatocellular carcinoma.

Figure 2

HMBOX1 inhibits HCC cell
proliferation, migration and colony formation in vitro. (A
and B) The effect of HMBOX1 overexpression in HCC cells
proliferation was assessed through a CCK-8 assay. (C and D) The
effect of HMBOX1 overexpression in HCC cells migration capacity was
assessed through the wound healing assay. (E) The effect of HMBOX1
overexpression in Huh-7 cell proliferation was assessed through the
colony formation assay. (F) The effect of HMBOX1 knockdown in Huh-7
cells proliferation was assessed through the CCK-8 assay. (G) The
effect of HMBOX1 knockdown in Huh-7 cells migration capacity was
assessed through the wound healing assay. *P<0.05 and
**P<0.01. HMBOX1, homeobox containing 1; HCC, hepatocellular
carcinoma; CCK-8, Cell Counting Kit-8; si-, small interfering; NC,
negative control.

Figure 3

Inhibitory effect of HMBOX1 on HCC is
dependent on the blocking of AKT1 phosphorylation. (A and B) The
difference in protein expression between HMBOX1-overexpressing
cells and control cells was analyzed through proteomics, and the
results are shown in the form of volcano map (A) and heat map (B)
respectively. (C and D) HCC cells were transfected with lentiviral
vector to overexpress HMBOX1 and then treated with 4 µM SC79 for
different times. The expression levels of AKT1 and p-AKT1 (Ser473)
were determined through western blotting in Hepa1-6 and Huh-7 cells
from different groups. (E-H) The proliferation and cell migration
ability of Hepa1-6 and Huh-7 cells from each group were analyzed
using a Cell Counting Kit-8 assay (E and F) and wound healing assay
(G and H) (I) The proliferation ability of Huh-7 cells from
different groups was assessed through the colony formation assay.
*P<0.05 and **P<0.01. HMBOX1, homeobox containing 1; HCC,
hepatocellular carcinoma.

Figure 4

HMBOX1 overexpression promotes PTPN1
expression in HCC cells. (A) The correlation between the expression
of HMBOX1 and PTPN1 in HCC tissues was analyzed via the GEPIA2
database. (B) IHC analysis of PTPN1 expression in HCC tumor tissues
and adjacent non-tumor tissues from 48 patients with HCC. (C) PTPN1
expression level was positively correlated with HMBOX1 expression
level in HCC tissues according to IHC analysis. (D) Effect of
overexpression of HMBOX1 on PTPN1 expression was determined through
western blotting. *P<0.05 and **P<0.01. HMBOX1, homeobox
containing 1; PTPN1, protein tyrosine phosphatase non-receptor type
1; HCC, hepatocellular carcinoma; IHC, immunohistochemical.

Figure 5

HMBOX1 inhibits HCC proliferation
ability by regulating PTPN1/AKT1 signaling. (A and B)
siRNA-targeted PTPN1 was used to silence PTPN1 in HCC cells which
overexpressing HMBOX1, and western blotting was used to detect the
expression of related molecules. (C and D) The proliferative
ability of HCC cells from each group was analyzed using the Cell
Counting Kit-8 assay. (E and F) Cell migration ability of HCC cells
from each group was measured through the wound healing assay.
*P<0.05, **P<0.01 and ***P<0.01. HMBOX1, homeobox
containing 1; PTPN1, protein tyrosine phosphatase non-receptor type
1; HCC, hepatocellular carcinoma; si-, small interfering; NC,
negative control.

Figure 6

Overexpression of HMBOX1 can inhibit
tumor growth in hepatocellular carcinoma bearing mice. A total of
5×106 Hepa1-6 cells overexpressing HMBOX1 (Over-HMBOX1)
or transfected with empty vector (Vector) were subcutaneously
injected into the right subaxillary region of C57BL/6 mice (n=6).
(A) Tumor growth curves were measured every three days. (B)
Representative images and the tumor weight of the subcutaneous
tumors from each group. (C) The expression levels of indicated
molecules were determined through western blotting in different
tumor tissues. **P<0.01. HMBOX1, homeobox containing 1; PTPN1,
protein tyrosine phosphatase non-receptor type 1; p-,
phosphorylated.
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Copy and paste a formatted citation
Spandidos Publications style
Zhang C, Jiang Y, Liu J, Zhang W, Qi J, Wen Q and Zhao H: <p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>. Oncol Rep 55: 47, 2026.
APA
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., & Zhao, H. (2026). <p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>. Oncology Reports, 55, 47. https://doi.org/10.3892/or.2026.9052
MLA
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., Zhao, H."<p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>". Oncology Reports 55.3 (2026): 47.
Chicago
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., Zhao, H."<p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>". Oncology Reports 55, no. 3 (2026): 47. https://doi.org/10.3892/or.2026.9052
Copy and paste a formatted citation
x
Spandidos Publications style
Zhang C, Jiang Y, Liu J, Zhang W, Qi J, Wen Q and Zhao H: <p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>. Oncol Rep 55: 47, 2026.
APA
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., & Zhao, H. (2026). <p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>. Oncology Reports, 55, 47. https://doi.org/10.3892/or.2026.9052
MLA
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., Zhao, H."<p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>". Oncology Reports 55.3 (2026): 47.
Chicago
Zhang, C., Jiang, Y., Liu, J., Zhang, W., Qi, J., Wen, Q., Zhao, H."<p>HMBOX1 inhibits hepatocellular carcinoma progression via PTPN1 mediated AKT1 phosphorylation</p>". Oncology Reports 55, no. 3 (2026): 47. https://doi.org/10.3892/or.2026.9052
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