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CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma

  • Authors:
    • Feng Li
    • Qing Dong
    • Zhe Kai
    • Qi Pan
    • Chunsheng Liu
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, The First People's Hospital of Anqing, Anqing, Anhui 241002, P.R. China, Department of General Surgery, The First People's Hospital of Anqing, Anqing, Anhui 241002, P.R. China, Department of Rehabilitation Medicine, The Sixth People's Hospital of Anqing, Anqing, Anhui 241002, P.R. China, Department of Laboratory Medicine, Yijishan Hospital of Wannan Medical College, Wuhu, Anhui 241001, PR. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 104
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    Published online on: June 27, 2025
       https://doi.org/10.3892/or.2025.8937
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Abstract

Cytochrome P450 8B1 (CYP8B1) is a critical enzyme in bile acid metabolism. Using multiple databases, including Gene Expression Omnibus, UALCAN (The University of Alabama at Birmingham Cancer data analysis Portal, GEPIA (Gene Expression Profiling Interactive Analysis), TCGA (The Cancer Genome Atlas) and GTEx (Genotype‑Tissue Expression), the present study analyzed CYP8B1 expression and its prognostic value in hepatocellular carcinoma (HCC). The results showed that CYP8B1 expression was significantly lower in HCC compared with normal tissues, and reduced CYP8B1 expression was associated with poor prognosis in patients with HCC. CYP8B1 was overexpressed in HCC cell lines (Huh7 and Hep3b cells); cell proliferation was assessed using Cell Counting Kit‑8 and EdU assays, while apoptosis was evaluated using the TUNEL assay. CYP8B1 overexpression inhibited proliferation and promoted apoptosis in HCC cells. Additionally, analyses via UALCAN and the Metascape platform showed that CYP8B1 expression was negatively associated with YWHAZ (Tyrosine 3/tryptophan 5 monooxygenase activation protein ζ), the regulation of PLK (Polo‑like kinase) activity during G2/M transition, and the intrinsic apoptosis pathway. Immunoblotting revealed that CYP8B1 overexpression decreased YWHAZ levels. Consistently, the expression of cyclin‑dependent kinase 1) and CCNB1 (Cyclin B1), key markers of G2/M transition, was also diminished following CYP8B1 overexpression. Furthermore, the pro‑apoptotic protein Bax was upregulated, while the anti‑apoptotic protein Bcl‑2 was downregulated. In conclusion, CYP8B1 holds promise as a potential prognostic target for HCC.
View Figures

Figure 1

CYP8B1 expression in public database
and clinical samples in LIHC. (A) Venn plots of (A) down- and (B)
upregulated overlapping DEGs. (C) Fold-change of overlapping DEGs.
CYP8B1 expression was significantly lower in tumor tissues compared
with normal tissue in LIHC in (D) University of Alabama at
Birmingham Cancer data analysis Portal), (E) Gene Expression
Profiling Interactive Analysis) and (F) TCGA and GTEx
(Genotype-Tissue Expression) database. CYP8B1 expression was
significantly lower in tumor compared with corresponding adjacent
normal tissue in (G) GSE84402 and (H) patients with hepatocellular
carcinoma. (I) Relative protein expression. *P<0.05,
**P<0.01. CYP8B1, cytochrome P450 8B1; LIHC, Liver
hepatocellular carcinoma; DEG, Differentially expressed gene; TCGA,
The Cancer Genome Atlas; THRSP, Thyroid hormone-responsive protein;
VIPR1, Vasoactive intestinal peptide receptor 1; MFSD2A, major
facilitator super family domain containing 2a; CDK,
Cyclin-dependent kinases; TPM, Transcripts per million; P,
patient.

Figure 2

Association between the OS and CYP8B1
expression in various LIHC clinical subgroups. Effect of CYP8B1 on
OS in patients with LIHC in (A) UALCAN (The University of Alabama
at Birmingham Cancer data analysis Portal) and (B) GEPIA (Gene
Expression Profiling Interactive Analysis)platform. Effect of
CYP8B1 on OS in patients with LIHC and (C) AFP (Alpha-fetoprotein)
≤400 ng/ml, (D) age ≤60 years, (E) male sex, (F) height ≥170 cm,
(G) Asian ethnicity and (H) residual tumor, R0. OS, Overall
survival; CYP8B1, Cytochrome P450 8B1; LIHC, Liver hepatocellular
carcinoma; TPM, Transcripts per million. A, low expression group
means the expression value <25%, medium expression group means
the value was 25–75%, and the high expression group means the value
was >75%. B-H, Arrange the CYP8B1 expression value from small to
larger, the low expression group means the expression value
<50%, and the high expression group means the expression
>50%.

Figure 3

Correlation analysis of CYP8B1
expression and immune cell infiltration. (A) Correlation analysis
of CYP8B1 expression and immune cell infiltration. Top three
positively correlated immune cells, (B) Th17 cells, (C) neutrophils
and (D) Tcm cells. Top three negatively correlated immune cells,
(E) Th2, (F) TFH and (G) NK CD56 bright cells. (H) CYP8B1
expression. CYP8B1, Cytochrome P450 8B1; Th, T helper cells; Tcm,
Central memory T cell; TFH, Follicular helper T cell; NK, Natural
killer; Treg, regulatory T cell; pDC, Plasmacytoid Dendritic cells;
Tgd, T gamma delta; aDC, active Dendritic cells; iDC,
interdigitating Dendritic cells; Tem, Effective memory T cell; Cor,
Correlation; TPM, Transcripts per million; ns, not significant.
*P<0.05, **P<0.01, ***P<0.001.

Figure 4

Efficiency of oe-CYP8B1 in HCC cell
lines. (A) mRNA and (B) protein expression level of CYP8B1 was
significantly increased in oe-CYP8B1 compared with NC in Huh7
cells. (C) mRNA and (D) protein expression of CYP8B1 was
significantly increased in oe-CYP8B1 compared with NC group in
Hep3b cells. **P<0.01. oe-CYP8B1, overexpressed Cytochrome P450
8B1; NC, Negative control .

Figure 5

o e-CYP8B1 inhibits proliferation of
hepatocellular carcinoma cells. (A) CCK-8 and (B) EdU assay showed
that oe-CYP8B1 (C) inhibited the proliferation of Huh7 cells. (D)
CCK-8 and (E) EdU assay showed that oe-CYP8B1 (F) inhibited
proliferation of Hep3b cells. *P<0.05, **P<0.01 vs. NC. Scale
bar, 100 µm. oe-CYP8B1, overexpressed cytochrome P450 8B1; CCK,
Cell counting kit; NC, Negative control; OD, optical density.

Figure 6

oe-CYP8B1 promotes the apoptosis of
hepatocellular carcinoma cells. (A) TUNEL assay indicated that
oe-CYP8B1 (B) promoted the apoptosis of Huh7 cells. (C) TUNEL assay
indicated that oe-CYP8B1 (D) promoted the apoptosis of Hep3b cells.
**P<0.01. Scale bar, 100 µm. The white arrows indicate
TUNEL-positive cells. oe-CYP8B1, overexpressed Cytochrome P450 8B1;
NC, Negative control.

Figure 7

Pathway and downstream biomarker
validation of CYP8B1 regulating HCC. Heatmap of the top 24 genes
(A) negatively and (B) positively correlated with CYP8B1 in LIHC in
UALCAN (The University of Alabama at Birmingham Cancer data
analysis Portal) platform. Enrichment analysis of genes (C)
negatively and (D) positively correlated with CYP8B1 in LIHC in
Metascape website. (E) Western blot assay of (F) biomarkers
downstream of CYP8B1 in HCC cells. *P<0.05, **P<0.01. CYP8B1,
cytochrome P450 8B1; LIHC, Liver hepatocellular carcinoma; HCC,
Hepatocellular carcinoma; NC, Negative control; oe, overexpressed;
YWHAZ, Tyrosine 3/tryptophan 5 monooxygenase activation protein
zeta; CDK, Cyclin-dependent kinases; TPM, Transcripts per
million.
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Copy and paste a formatted citation
Spandidos Publications style
Li F, Dong Q, Kai Z, Pan Q and Liu C: CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma. Oncol Rep 54: 104, 2025.
APA
Li, F., Dong, Q., Kai, Z., Pan, Q., & Liu, C. (2025). CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma. Oncology Reports, 54, 104. https://doi.org/10.3892/or.2025.8937
MLA
Li, F., Dong, Q., Kai, Z., Pan, Q., Liu, C."CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma". Oncology Reports 54.3 (2025): 104.
Chicago
Li, F., Dong, Q., Kai, Z., Pan, Q., Liu, C."CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma". Oncology Reports 54, no. 3 (2025): 104. https://doi.org/10.3892/or.2025.8937
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Spandidos Publications style
Li F, Dong Q, Kai Z, Pan Q and Liu C: CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma. Oncol Rep 54: 104, 2025.
APA
Li, F., Dong, Q., Kai, Z., Pan, Q., & Liu, C. (2025). CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma. Oncology Reports, 54, 104. https://doi.org/10.3892/or.2025.8937
MLA
Li, F., Dong, Q., Kai, Z., Pan, Q., Liu, C."CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma". Oncology Reports 54.3 (2025): 104.
Chicago
Li, F., Dong, Q., Kai, Z., Pan, Q., Liu, C."CYP8B1 is a prognostic biomarker with important functional implications in hepatocellular carcinoma". Oncology Reports 54, no. 3 (2025): 104. https://doi.org/10.3892/or.2025.8937
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