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PSMD6 as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and in vitro validation

  • Authors:
    • Jian Liu
    • Ting Wang
    • Zhenhua Zhu
  • View Affiliations / Copyright

    Affiliations: Department of Emergency, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, P.R. China, Department of Public Health, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, P.R. China, Otolaryngology‑Wide Head and Neck Surgery Department, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 28
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    Published online on: February 24, 2026
       https://doi.org/10.3892/mco.2026.2937
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Abstract

Hepatocellular carcinoma (HCC) is a lethal malignancy with limited diagnostic biomarkers. The present study aims to comprehensively investigate the expression, clinical prognostic significance, and potential mechanisms of PSMD6 in HCC through comprehensive bioinformatics analyses and in vitro experimental validation. PSMD6 expression in HCC was analyzed using data from the UALCAN, SANGERBOX and TIMER databases. The association between PSMD6 expression and clinicopathological features, patient prognosis, and immune cell filtration was evaluated. Functional enrichment analyses (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes) were performed to identify PSMD6‑related signaling pathways, and a protein‑protein interaction (PPI) network was constructed using the BioGRID and STRING databases. Key findings from bioinformatics analyses were validated in vitro using reverse transcription‑quantitative (RT‑qPCR) and western blotting in HCC cell lines. PSMD6 expression was significantly upregulated in HCC compared with adjacent normal tissues (P<0.001), which has been consistently validated by various databases and confirmed by in vitro experiments using RT‑qPCR and western blotting. High PSMD6 expression was significantly associated with advanced tumor grade and patient age and served as an independent predictor of poor overall survival (P<0.001). In addition, PSMD6 demonstrated high diagnostic accuracy for HCC (area under the curve=0.877). Moreover, PSMD6 expression showed a positive correlation with the infiltration levels of CD4+ T cells and B cells in HCC (P<0.05), independent of tumor purity (P>0.05). Functional enrichment analysis indicated that PSMD6 was involved in critical oncogenic pathways, including the cell cycle. PPI network analysis revealed that PSMD6 interact with several key proteins, such as PSMC3, PSMD7 and UCHL5 to achieve a regulatory function in HCC. In conclusion, PSMD6 is significantly overexpressed in HCC and is strongly associated with tumor progression and poor prognosis. It represents a promising diagnostic biomarker and a potential therapeutic target for HCC.
View Figures

Figure 1

Pan-cancer analysis of PSMD6
expression. (A) PSMD6 mRNA expression levels across various cancer
types from the TCGA project, as analyzed via the SANGERBOX
database. (B) Validation of PSMD6 expression in multiple cancers
using the UALCAN database. Data are presented as log2-transformed
TPM values. PSMD6 is significantly upregulated in LIHC compared
with normal tissue *P<0.05, **P<0.01,
***P<0.001 and ****P<0.0001. PSMD6,
proteasome 26s subunit, non-ATPase 6; TCGA, The Cancer Genome
Atlas; TPM, transcripts per million; LIHC, liver hepatocellular
carcinoma.

Figure 2

Differential expression of PSMD6 in
LIHC. (A and B) PSMD6 expression is significantly higher in LIHC
tumor tissues (n=371) compared with (A) adjacent normal tissues
(n=50) from TCGA or (B) combined normal tissues from TCGA and GTEx
(n=163), as analyzed by XIANTAO tools and UALCAN, respectively.
***P<0.001. PSMD6, proteasome 26s subunit, non-ATPase
6; LIHC, liver hepatocellular carcinoma; TCGA, The Cancer Genome
Atlas.

Figure 3

Association between PSMD6 expression
and clinicopathological characteristics in LIHC. (A-I) Box plots
showing PSMD6 expression levels stratified by (A) pathologic stage,
(B) tumor status, (C) ethnicity, (D) patient weight, (E) pathologic
T stage, (F) sex, (G) pathologic M stage, (H) age and (I)
pathologic N stage. Analyses were performed using data from The
Cancer Genome Atlas-LIHC cohort via XIANTAO tools.
*P<0.05, **P<0.01 and
***P<0.001. PSMD6, proteasome 26s subunit, non-ATPase
6; LIHC, liver hepatocellular carcinoma.

Figure 4

Survival analysis of PSMD6 in LIHC
using UALCAN. Kaplan-Meier curves showing the impact of PSMD6
expression on patient survival. (A) OS of all patients with LIHC
stratified by high and low PSMD6 expression. (B-D) Subgroup
analysis of OS based on (B) tumor grade, (C) ethnicity and (D) sex.
PSMD6, proteasome 26s subunit, non-ATPase 6; LIHC, liver
hepatocellular carcinoma; OS, overall survival.

Figure 5

Prognostic value of PSMD6 in LIHC
using SANGERBOX. (A-D) Kaplan-Meier survival analysis demonstrating
the relationship between PSMD6 expression and (A) overall survival,
(B) progression-free interval, (C) disease-free interval, and (D)
disease-specific survival. PSMD6, proteasome 26s subunit,
non-ATPase 6; HR, hazard ratio.

Figure 6

Diagnostic and prognostic value of
PSMD6. (A) Kaplan-Meier plot from the Kaplan-Meier Plotter database
confirms that high PSMD6 expression predicts poor overall survival
in patients with LIHC. (B) Receiver operating characteristic curve
analysis (via XIANTAO tools) evaluating the diagnostic power of
PSMD6 to distinguish LIHC tumors from normal tissues (FDR=1%).
PSMD6, proteasome 26s subunit, non-ATPase 6; LIHC, liver
hepatocellular carcinoma; AUC, area under the curve; HR, hazard
ratio; CI, confidence interval.

Figure 7

Validation of PSMD6 mRNA expression
in cell lines. Relative mRNA expression levels of PSMD6 in a normal
human hepatocyte cell line (THLE-2) and three hepatocellular
carcinoma cell lines (HepG2, MHCC97-L and MHCC97-H) as determined
by reverse transcription-quantitative PCR. Data are presented as
the mean ± SD. ***P<0.001 and
****P<0.0001. PSMD6, proteasome 26s subunit,
non-ATPase 6.

Figure 8

Validation of PSMD6 protein
expression in cell lines. Western blot analysis confirming the
elevated protein expression of PSMD6 in hepatocellular carcinoma
cell lines (HepG2, MHCC97-L and MHCC97-H) compared with the normal
hepatocyte cell line THLE-2. GAPDH was used as a loading control.
PSMD6, proteasome 26s subunit, non-ATPase 6.

Figure 9

Correlation between PSMD6 expression
and immune cell infiltration in the LIHC tumor microenvironment.
(A) Scatter plots showing the correlation between PSMD6 expression
(log2 TPM) and the abundance of various immune cell types,
estimated by the TIMER2.0 algorithm. Statistical significance was
determined by Spearman's correlation. (B) Overview of the TISCH
database analysis showing the immune landscape of LIHC. (C and D)
Single-cell RNA-seq analysis from the TISCH database illustrating
the distribution and expression levels of PSMD6 (color scale)
across immune cell clusters in the (C) GSE140228_Smartseq2 and (D)
GSE98638 datasets. PSMD6, proteasome 26s subunit, non-ATPase 6;
LIHC, liver hepatocellular carcinoma.

Figure 10

Functional enrichment analysis of
PSMD6-coexpressed genes in LIHC. (A) Significantly enriched Kyoto
Encyclopedia of Genes and Genomes pathways and (B) Gene Ontology
terms (Biological Process, Cellular Component, Molecular Function)
for genes correlated with PSMD6 expression in The Cancer Genome
Atlas-LIHC cohort. The bubble chart size and color represent the
number of genes and the significance level (-log10(P-value)),
respectively. PSMD6, proteasome 26s subunit, non-ATPase 6; LIHC,
liver hepatocellular carcinoma.

Figure 11

Analysis of the interaction between
proteasome 26s subunit, non-ATPase 6 and protein using BioGRID
database.

Figure 12

Analyzing the Interaction between
proteasome 26s subunit, non-ATPase 6 and proteins in STRING
database.
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Copy and paste a formatted citation
Spandidos Publications style
Liu J, Wang T and Zhu Z: <em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation. Mol Clin Oncol 24: 28, 2026.
APA
Liu, J., Wang, T., & Zhu, Z. (2026). <em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation. Molecular and Clinical Oncology, 24, 28. https://doi.org/10.3892/mco.2026.2937
MLA
Liu, J., Wang, T., Zhu, Z."<em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation". Molecular and Clinical Oncology 24.4 (2026): 28.
Chicago
Liu, J., Wang, T., Zhu, Z."<em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation". Molecular and Clinical Oncology 24, no. 4 (2026): 28. https://doi.org/10.3892/mco.2026.2937
Copy and paste a formatted citation
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Spandidos Publications style
Liu J, Wang T and Zhu Z: <em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation. Mol Clin Oncol 24: 28, 2026.
APA
Liu, J., Wang, T., & Zhu, Z. (2026). <em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation. Molecular and Clinical Oncology, 24, 28. https://doi.org/10.3892/mco.2026.2937
MLA
Liu, J., Wang, T., Zhu, Z."<em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation". Molecular and Clinical Oncology 24.4 (2026): 28.
Chicago
Liu, J., Wang, T., Zhu, Z."<em>PSMD6</em> as a biomarker for hepatocellular carcinoma: A comprehensive bioinformatics analysis and <em>in vitro</em> validation". Molecular and Clinical Oncology 24, no. 4 (2026): 28. https://doi.org/10.3892/mco.2026.2937
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