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Article Open Access

FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer

  • Authors:
    • Jin Liu
    • Maoyuan Feng
    • Jian Zhou
    • Shijie Yang
    • Yue Shi
    • Wenping Li
  • View Affiliations / Copyright

    Affiliations: Department of Urology, Hebei Medical University Third Hospital, Shijiazhuang, Hebei 050051, P.R. China, China National Center for Bioinformation, Beijing 100101, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 570
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    Published online on: October 3, 2025
       https://doi.org/10.3892/ol.2025.15316
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Abstract

F‑box and leucine‑rich repeat protein 22 (FBXL22) has been implicated in breast and prostate cancer types; however, comprehensive pan‑cancer analyses and detailed investigations of its role in cancer development are still needed. To address this, the present study used The Cancer Genome Atlas, Genotype‑Tissue Expression, University of ALabama at Birmingham CANcer data analysis Portal, Kaplan‑Meier‑Plotter and cBioPortal databases to analyze the correlation between FBXL22 expression and prognosis, gene mutations, DNA methylation, immune cell infiltration and immune‑related gene regulation in several types of cancer. The findings of the present study demonstrated that FBXL22 was predominantly downregulated in several cancer types and may serve as a prognostic and diagnostic marker in certain cancer types, particularly prostate cancer. Strong correlations between FBXL22 expression and immune checkpoint genes were observed, which suggests a role in the tumor immune microenvironment. Additionally, FBXL22 expression was associated with infiltration of cancer‑associated fibroblasts and endothelial cells, which may affect immunotherapy outcomes. Mechanistically, FBXL22 was involved in circadian rhythm regulation, ubiquitin‑mediated proteolysis, focal adhesion signaling and cGMP‑protein kinase G signaling. In prostate cancer, FBXL22 upregulation suppressed cell viability, invasion and metastasis, while promoting apoptosis, potentially through modulation of the polo‑like kinase 1 pathway. To the best of our knowledge, the present study provides the first comprehensive pan‑cancer analysis of FBXL22, which highlights its potential as an immune biomarker and therapeutic target for multiple cancer types with implications for precision medicine.
View Figures

Figure 1

Expression levels of FBXL22 in
pan-cancer. (A) The expression status of FBXL22 in different cancer
tissues and para-cancerous tissues in TCGA database. Error bars
represent standard deviation. (B) Expression status of FBXL22 in
matched cancer tissues and normal tissues in TCGA and GTEx
databases. Cancer names in red text indicate high FBXL22 expression
in this cancer, while blue text indicates low FBXL22 expression in
this cancer. (C) Correlation between FBXL22 expression and
methylation in TCGA database (D) Promoter methylation levels of
FBXL22 in PRAD samples. (E) Expression differences of FBXL22 in
PRAD samples based on sample types. *P<0.05; **P<0.01;
***P<0.001; ns, P≥0.05. FBXL22, F-box and leucine rich repeat
protein 22; ACC, adrenocortical carcinoma; BLCA, bladder urothelial
carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous
cell carcinoma and endocervical adenocarcinoma; CHOL,
cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid
neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma;
GBM, glioblastoma multiforme; HNSC, head and neck squamous cell
carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell
carcinoma; KIRP, kidney renal papillary cell carcinoma; LAML, acute
myeloid leukemia; LGG, brain lower grade glioma; LIHC, Liver
hepatocellular carcinoma; LUAD, Lung adenocarcinoma; LUSC, lung
squamous cell carcinoma; MESO, mesothelioma; ns, not significant;
OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic
adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD,
prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC,
sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach
adenocarcinoma; TCGA, The Cancer Genome Atlas; TGCT, testicular
germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; TMB,
tumor mutation burden; TPM, transcripts per million; UCEC, uterine
corpus endometrial carcinoma; UCS, uterine carcinosarcoma.

Figure 2

Clinical diagnosis and prognostic
implications of FBXL22 in PCa. (A) Kaplan-Meier curve of PFI in
patients with intermediate to advanced disease stage (Clinical T
stage, T2, T3 and T4) in PCa by FBXL22-expression groups. (B)
Diagnostic ROC curves of FBXL22 and other genes such as SPOP,
HOXB13, CHEK2, TP53, BRCA2 and BRCA1 in PCa. (C) FBXL22 expression
across PCa disease course subtypes. (D) FBXL22 expression in PCS
subtypes. (E) FBXL22 expression in basal, LumA and LumB subtypes.
(F) FBXL22 expression across GS subgroups. (G) FBXL22 expression in
PCS subtypes. (H) FBXL22 expression in basal, LumA and LumB
subtypes. FBXL22, F-box and leucine rich repeat protein 22; PCa,
prostate cancer; PFI, progression-free interval; ROC, receiver
operating characteristic; SPOP, Speckle-type pox virus and zinc
finger protein; HOXB13, homeobox B13; CHEK2, checkpoint kinase 2;
PAM50, Prediction Analysis of Microarray 50; PCTA, Prostate Cancer
Transcriptome Atlas; TCGA, The Cancer Genome Atlas; PCS, PCa
subtypes; ROC, receiver operating characteristic; Lum, luminal; GS,
Gleason score; TPR, true positive rate; FPR, false positive rate;
AUC, area under the curve.

Figure 3

Association of FBXL22 with
immune-infiltration in PCa. (A) The immune score of
FBXL22-expression groups. (B) Expression levels of the 17 immune
checkpoint proteins by FBXL22-expression group. (C) Heatmap
illustrating correlations between FBXL22 expression and 17 immune
checkpoint proteins. (D) Correlation between FBXL22 expression and
the infiltration levels of various immune cells. Scatter plot of
correlation between FBXL22 expression and the infiltration levels
of (E) NK, (F) Tgd, (G) Th1 and (H) DC cells. (I) Infiltration
levels of 23 types of immune cells were assessed by
FBXL22-expression group using ssGSEA. *P<0.05, **P<0.01 and
***P<0.001; ns, P≥0.05. FBXL22, F-box and leucine rich repeat
protein 22; PCa, prostate cancer; NK, natural killer; DC, dendritic
cells; Tgd cells, γδ T cells; Th1 cells, T helper type 1 cells;
Treg, regulatory T cells; ssGSEA, single-sample gene set enrichment
analysis; ESTIMATE, Estimation of STromal and Immune cells in
MAlignant Tumor tissues using Expression data; TPM, transcripts per
million; ns, not significant.

Figure 4

Predictive effects of therapeutic
response for FBXL22 in pan-cancer. Kaplan-Meier curves of patients
treated with immunotherapy by FBXL22 expression for (A) OS and (B)
PFS. Kaplan-Meier curves for (C) OS and (D) PFS of patients treated
with atezolizumab by FBXL22 expression. Correlation between FBXL22
expression and (E) MATH, (F) TMB, (G) MSI and (H) HRD. FBXL22,
F-box and leucine rich repeat protein 22; MATH, mutant allele tumor
heterogeneity; TMB, tumor mutation burden; MSI, microsatellite
instability; HRD, homologous recombination deficiency; OS, overall
survival; PFS; progression-free survival; HR, hazard ratio.

Figure 5

Correlation analysis between FBXL22
expression and immune infiltration of CAFs and ECs across all types
of cancer in TCGA. (A) Corresponding heatmap and (B) COAD, (C) LGG,
(D) TGCT, (E) THCA, (F) PRAD, (G) LGG, (H) PAAD, (I) PRAD, (J)
THCA, (K) READ. Each scatter plot includes data points representing
FBXL22 expression levels (x-axis) and infiltration levels of CAFs
or ECs (y-axis), with a trend line indicating the correlation
direction. Spearman's correlation coefficient (ρ) and corresponding
P-value are annotated for each plot to quantify the strength and
significance of the association. FBXL22, F-box and leucine rich
repeat protein 22; CAFs, cancer-associated fibroblasts; ECs,
endothelial cells; TCGA, The Cancer Genome Atlas; TPM, transcripts
per million; COAD, colon adenocarcinoma; PRAD, prostate
adenocarcinoma; READ, rectum adenocarcinoma; TGCT, testicular germ
cell tumors; THCA, thyroid carcinoma; LGG, lower grade glioma.

Figure 6

FBXL22-related gene enrichment
analysis in pan-cancer. (A) Network of interactions among
FBXL22-binding proteins using the STRING tool. (B) Heatmap of
expression correlations between FBXL22 and the top 10
FBXL22-correlated genes in different cancer types of TCGA using the
GEPIA2 tools, including MRGPRF, KCNMB1, PDLIM7, TAGLN, TGFB1I1,
JPH2, CNN1, FLNA, CSRP1 and MYL9. (C) KEGG and GO enrichment
analyses are visualized by cluster tree diagram. FBXL22, F-box and
leucine rich repeat protein 22; STRING, Search Tool for the
Retrieval of Interacting Genes/Proteins; TCGA, The Cancer Genome
Atlas; GEPIA2, Gene Expression Profiling Interactive Analysis 2;
MRGPRF, Mas-related G protein-coupled receptor F; KCNMB1, potassium
calcium-activated channel subfamily M regulatory β subunit 1;
PDLIM7L, PDZ and LIM domain 7; TAGLN, Transgelin; TGFB1I1,
transforming growth factor β 1 induced 1; JPH2, junctophilin 2;
CNN1, calponin 1; FLNA, filamin A; CSRP1, cysteine and glycine rich
protein 1; MYL9, myosin light chain 9; KEGG, Kyoto Encyclopedia of
Genes and Genomes; GO, Gene ontology; BP, Biological process; CC,
Cellular component; MF, Molecular function.

Figure 7

Differential expression and
functional enrichment analysis of FBXL22-associated genes in
prostate cancer. (A) Volcano plot illustrating differential
expression analysis between high-FBXL22 and low-FBXL22 expression
groups in PCa, which highlights the genes that were significantly
upregulated or downregulated in association with FBXL22 expression
levels. The results of KEGG and GO enrichment analysis are depicted
using (B) heat map and (C) Enrichment Map Analysis Platform,
respectively. (D) The analysis of GO/KEGG combined log(fold change)
is represented by a chordal diagram. The mountain maps display the
pathways (E) significantly negative or (F) significantly positive
associated with expression levels of FBXL22 in GSEA analysis
results. Error bars represent standard deviation. FBXL22, F-box and
leucine rich repeat protein 22; PCa, prostate cancer; KEGG, Kyoto
Encyclopedia of Genes and Genomes; GO, Gene Ontology; GSEA, Gene
Set Enrichment Analysis; E2F, E2 DNA-binding factor; PLK1,
polo-like kinase 1; NES, nuclear export sequence; FDR, false
discovery rate; BP, biological process; CC, cellular component; MF,
molecular function.

Figure 8

Effects of FBXL22 on viability,
invasion, migration and apoptosis of PCa cells. (A) The relative
expression levels of FBXL22 in normal prostate epithelial cell
lines RWPE-1 and PCa cell lines PC3 and DU145. The relative
expression levels of FBXL22 in NC and FBXL22-OE in (B) PC3 and (C)
DU145 cell lines. Semi-quantification of relative protein levels of
FBXL22 in NC and FBXL22-OE groups in (D) PC3 and (E) DU145 cells.
Western blot of FBXL22 in NC and FBXL22-OE groups in (F) PC3 and
(G) DU145 cells. Cell viability was assessed using the CCK-8 assay
in (H) PC3 and (I) DU145 cells. (J-M) The Transwell chamber was
utilized to evaluate cell invasive ability (scale bar, 50 µm).
(N-Q) Cell migratory ability was examined using a wound healing
assay (scale bar, 250 µm). The histograms demonstrate the relative
percentage of wound healing. (R) Quantification of apoptosis rates
in PC3 cells transfected with FBXL22-OE compared with the NC group,
shown as a bar graph. (S) Quantification of apoptosis rates in
DU145 cells under the same conditions. (T) Representative flow
cytometry dot plots for PC3 cells, detailing the distribution of
cells in different apoptosis stages. (U) Representative flow
cytometry dot plots for DU145 cells, using annexin V-FITC and PI
staining to delineate cell viability and apoptosis stages. NK cell
migration and cytotoxicity against FBXL22-OE (V) PC3 and (W) DU145
cells in co-culture Transwell experiments (scale bar, 50 µm).
Co-immunoprecipitation analysis confirming the interaction between
FBXL22 and PLK1 in (X) PC3 and (Y) DU145 cells. Error bars
represent standard deviation. **P<0.01, ***P<0.001 and
****P<0.0001; ns, P≥0.05. PCa, prostate cancer; FBXL22, F-box
and leucine rich repeat protein 22; OE, overexpression; NC,
negative control; ns, not significant; NK, natural killer.

Figure 9

Possible mechanism underlying the
impact of FBXL22 on PCa cell function. Western blot analysis
demonstrated the relative levels of p-PLK1 and PLK1 in (A) PC3 and
(B) DU145 cells transfected with FBXL22-OE compared with the NC
group. (C) Representative western blot images for p-PLK1, PLK1 and
GAPDH in PC3 and DU145 cells. Western blot analysis indicated the
relative levels of AR-V7 in (D) PC3 and (E) DU145 cells. (F)
Representative western blot images for AR-V7, PD-L1 and GAPDH in
PC3 and DU145 cells. Western blot analysis indicated the relative
levels of PD-L1 in (G) PC3 and (H) DU145 cells. (I) Representative
western blot images for cleaved-caspase-3, caspase-3,
cleaved-caspase-9, caspase-9, caspase-8 and GAPDH in PC3 and DU145
cells. (J) Mechanism by which FBXL22 functions in PCa cells. Error
bars represent standard deviation. The data indicates that FBXL22
overexpression reduces p-PLK1, AR-V7 and PD-L1 levels while
increasing cleaved caspase-3, −8 and −9 levels, which suggests
modulation of the PLK1 pathway and promotion of apoptosis.
**P<0.01, ***P<0.001 and ****P<0.0001. PCa, prostate
cancer; FBXL22, F-box and leucine rich repeat protein 22; OE,
overexpression; NC, negative control; PLK1, polo-like kinase 1;
AR-V7, androgen receptor splice variant 7; PD-L1, programmed
death-ligand 1; PDCD1, programmed cell death 1; PDCDILG2,
programmed cell death 1 Ligand 2.
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Copy and paste a formatted citation
Spandidos Publications style
Liu J, Feng M, Zhou J, Yang S, Shi Y and Li W: FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer. Oncol Lett 30: 570, 2025.
APA
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., & Li, W. (2025). FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer. Oncology Letters, 30, 570. https://doi.org/10.3892/ol.2025.15316
MLA
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., Li, W."FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer". Oncology Letters 30.6 (2025): 570.
Chicago
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., Li, W."FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer". Oncology Letters 30, no. 6 (2025): 570. https://doi.org/10.3892/ol.2025.15316
Copy and paste a formatted citation
x
Spandidos Publications style
Liu J, Feng M, Zhou J, Yang S, Shi Y and Li W: FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer. Oncol Lett 30: 570, 2025.
APA
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., & Li, W. (2025). FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer. Oncology Letters, 30, 570. https://doi.org/10.3892/ol.2025.15316
MLA
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., Li, W."FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer". Oncology Letters 30.6 (2025): 570.
Chicago
Liu, J., Feng, M., Zhou, J., Yang, S., Shi, Y., Li, W."FBXL22, a circadian‑regulated immune biomarker with pan‑cancer prognostic value and therapeutic potential in prostate cancer". Oncology Letters 30, no. 6 (2025): 570. https://doi.org/10.3892/ol.2025.15316
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