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CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling

  • Authors:
    • Yu Wang
    • Huandi Zhou
    • Xuetao Han
    • Dongdong Zhang
    • Liubing Hou
    • Haonan Li
    • Tianyi Fan
    • Sijie Li
    • Xiaoying Xue
  • View Affiliations / Copyright

    Affiliations: Department of Radiotherapy, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, P.R. China
    Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 41
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    Published online on: January 12, 2026
       https://doi.org/10.3892/or.2026.9046
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Abstract

Glioblastoma remains a lethal malignancy with limited therapeutic advancements. Emerging evidence implicates cell cycle dysregulation in glioma pathogenesis, yet the mechanistic role of cyclin‑dependent kinase 1 (CDK1) remains underexplored. The present study systematically evaluated the clinical relevance and functional impact of CDK1 in glioma progression through multi‑modal experimental approaches. CDK1 expression was analyzed using public datasets and then verified by western blotting using patient tissue samples (n=37) from the Second Hospital of Hebei Medical University (Shijiazhuang, China). Survival analysis was performed using Chinese Glioma Genome Atlas and The Cancer Genome Atlas datasets, alongside multivariate Cox regression to evaluate prognostic independence. Functional assays, including small interfering RNA‑mediated CDK1 knockdown, were conducted in glioma cell lines to assess proliferation (Cell Counting Kit‑8 and EdU), migration/invasion (Transwell), apoptosis (acridine orange/ethidium bromide staining and flow cytometry) and radiosensitivity (γ‑H2AX foci quantification post‑irradiation). The expression levels of downstream cell cycle regulators were quantified via quantitative PCR. The results indicated that CDK1 was significantly upregulated in glioma tissues compared with normal controls, with expression levels escalating with tumor grade. High CDK1 expression correlated with a reduced overall survival and served as an independent prognostic marker. CDK1 knockdown attenuated glioma cell proliferation, migration and invasion, while enhancing apoptosis and radiosensitivity. Mechanistically, CDK1 knockdown downregulated cell cycle regulators proliferating cell nuclear antigen, minichromosome maintenance complex component 2‑4 (MCM2‑4), MCM6, polo‑like kinase 1, TTK protein kinase and mitotic arrest deficient 2 like 1, implicating mitotic dysregulation as a central pathway. The present study established CDK1 as a master regulator of glioma progression through coordinated control of proliferation, DNA repair and metastatic potential. The robust association between CDK1 expression, tumor grade and survival, coupled with functional validation across complementary assays, positions CDK1 inhibition as a promising therapeutic strategy. The mechanistic elucidation of its cell cycle network provides a novel framework for targeting glioma‑specific therapeutic targets.

View Figures

Figure 1

Workflow of the whole study. FC, fold
change; DEGs, differentially expressed genes; CDK1,
cyclin-dependent kinase 1; GEPIA, Gene Expression Profiling
Interactive Analysis; N, normal tissue; T, tumor; CGGA, Chinese
Glioma Genome Atlas; TCGA, The Cancer Genome Atlas; ROC, receiver
operating characteristic; AUC, area under the curve; PRS,
prognostic risk score; IDH, isocitrate dehydrogenase; MGMT,
O-6-methylguanine-DNA methyltransferase; OS, overall survival; WHO,
World Health Organization; NC, negative control; si, small
interfering RNA; OD, optical density; AO/EB, acridine
orange/ethidium bromide; GSEA, Gene Set Enrichment Analysis; PPI,
protein-protein interaction; PCNA, proliferating cell nuclear
antigen; MCM2/3/4/6, minichromosome maintenance complex component
2/3/4/6; PLK1, polo-like kinase 1; MAD2L1, mitotic arrest deficient
2 like 1; GBM, glioblastoma; LGG, low-grade glioma.

Figure 2

Core gene and its expression levels
in normal tissue and glioma were retrieved. (A) Volcano plots of
the 5,715 DEGs in the GSE29796 dataset (599 upregulated and 5,116
downregulated). (B) Volcano plots of the 2,450 DEGs in the GSE4290
dataset (1,425 upregulated and 1,025 downregulated). (C) Volcano
plots of the 4,696 DEG in the GSE50161 dataset (2,320 upregulated
and 2,376 downregulated). (D) Volcano plots of the 1,532 DEGs in
the GSE7696 dataset (915 upregulated and 617 downregulated). The
red nodes represent the significantly upregulated genes with
log2FC>1 and P<0.05. The green nodes represent the
significantly downregulated genes with log2FC<-1 and P<0.05.
(E) Intersecting genes of the four groups of DEGs are displayed by
Venn diagram. (F) Protein-protein interaction network was
constructed and the core protein with the highest degree of
topology value was screened out, based on the STRING website and
Cytoscape visualization software. Comparison of CDK1 mRNA
expression levels in normal tissue and glioma using the (G)
GSE29796, (H) GSE4290, (I) GSE50161 and (J) GSE7696 datasets. (K)
CDK1 expression levels in normal tissue and glioma via GEPIA
analysis. (L) The protein levels of CDK1 in normal (intensity:
negative; quantity: none) and glioma (intensity: strong; quantity:
<25%) tissue from the Human Protein Atlas. (M) Western blot
assay and (N) Wilcoxon matched-pairs signed rank test (P<0.0001)
of 37 pairs of glioma and adjacent tissues. (O) Data from the
internal cohort showed that CDK1 expression was a statistically
significant difference between Grade 2+3 gliomas (n=20) and Grade 4
gliomas (n=17) (P=0.0018). Data are presented as mean ± SD.
*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
Statistical significance was calculated using unpaired t-test, Mann
Whitney test or Wilcoxon matched-pairs signed rank test. FC, fold
change; DEGs, differentially expressed genes; CDK1,
cyclin-dependent kinase 1; GEPIA, Gene Expression Profiling
Interactive Analysis; N, normal tissue; T, tumor.

Figure 3

Survival analysis and independent
prognostic analysis of CDK1 based on bioinformatics analysis.
Survival analysis of the high and low CDK1 groups in patients with
glioma from the (A) CGGA and (B) TCGA datasets. The ROC curves of
CDK1 at 1, 3 and 5 year survival in the (C) CGGA and (D) TCGA
datasets. Univariate analysis of CDK1 in the (E) CGGA and (F) TCGA
datasets. Multivariate analysis of CDK1 in the (G) CGGA and (H)
TCGA datasets. (I) Prognostic nomogram constructed based on the
CGGA dataset including clinical information to predict the survival
of patients with glioma. Calibration curves of the nomogram
constructed based on the CGGA dataset to predict survival at (J) 1
year, (K) 2 years and (L) 3 years. CDK1, cyclin-dependent kinase 1;
CGGA, Chinese Glioma Genome Atlas; TCGA, The Cancer Genome Atlas;
ROC, receiver operating characteristic; AUC, area under the curve;
PRS, prognostic risk score; IDH, isocitrate dehydrogenase; MGMT,
O-6-methylguanine-DNA methyltransferase; OS, overall survival.

Figure 4

Association between CDK1 and the
clinicopathological characteristics of patients with glioma in the
Chinese Glioma Genome Atlas dataset. (A) The CDK1 expression level
by age (≥42 vs. <42 years; P=0.002). (B) The CDK1 expression
level by glioma grade (WHO II, III and IV; P<0.001). (C) The
CDK1 expression level by 1p19q codeletion status (codel vs.
non-codel; P<0.001). (D) The CDK1 expression level by IDH
mutation state (mutant vs. wild-type; P<0.001). (E) The CDK1
expression level by glioma type (primary, recurrent and secondary;
P<0.001). (F) The CDK1 expression level in IDH-wildtype
glioblastoma vs. IDH-mutant astrocytoma and oligodendroglioma
(P<0.001). (G) The CDK1 expression level in IDH-mutant
astrocytoma (1p/19q non-codel) vs. IDH-mutant oligodendroglioma
(1p/19q code) (P=0.0305). Data are presented as mean ± SD.
Statistical significance was calculated using unpaired t-test or
Mann Whitney test. CDK1, cyclin-dependent kinase 1; WHO, World
Health Organization; IDH, isocitrate dehydrogenase.

Figure 5

Verification of CDK1 knockdown and
its role in the proliferative ability of glioma cells. Validation
of CDK1 knockdown was achieved through western blot analysis in the
(A) U251MG and (B) LN229 glioma cell lines. (C and D) Statistical
comparisons of the western blot data for the (C) U251MG (Blank vs.
NC, P=0.9992; Blank vs. CDK1-si, P=0.0057; NC vs. CDK1-si,
P=0.0053) and LN229 (Blank vs. NC, P=0.7960; Blank vs. CDK1-si,
P=0.0161; NC vs. CDK1-si, P=0.0343) cell lines (n=3, each group).
(E and F) Cell Counting Kit-8 cell viability assays demonstrated a
decrease in proliferation potential upon CDK1 knockdown in (E)
U251MG and (F) LN229 cells (n=4, each group). (G) Representative
images (magnification, ×10) and (H) quantitative analysis of the
EdU assay performed in the U251MG cell line (NC vs. CDK1-si,
P=0.0325; n=3, each group). (I) Representative images
(magnification, ×10) and (J) quantitative analysis of the EdU assay
performed in the LN229 cell line (NC vs. CDK1-si, P=0.0057; n=3,
each group). Data are presented as mean ± SD. Statistical
significance was calculated using one-way analysis of variance
followed by Tukey's multiple comparisons test, unpaired t-test or
one-way analysis of variance followed by Sidak's multiple
comparisons test. *P<0.05, **P<0.01, ***P<0.001. CDK1,
cyclin-dependent kinase 1; NC, negative control; si, small
interfering (RNA); OD, optical density.

Figure 6

Impact of CDK1 on the
migration/invasion capability and DNA damage repair ability of
glioma cells. (A) Representative images of the U251MG cell
Transwell migration assay (magnification, ×10) and the (B)
quantitative analysis of the number of migrated cells (NC vs.
CDK1-si, P<0.001; n=3). (C) Representative images of the LN229
cell Transwell migration assay (magnification, ×10) and the (D)
quantitative analysis of the number of migrated cells (NC vs.
CDK1-si, P=0.0002; n=3). (E) Representative images of the U251MG
cell Transwell invasion assay (magnification, ×10) and the (F)
quantitative analysis (NC vs. CDK1-si, P<0.001; n=3). (G)
Representative images of the LN229 cell Transwell invasion assay
(magnification, ×10) and the (H) quantitative analysis (NC vs.
CDK1-si, P=0.0003; n=3). (I) The assessment of cell migration via
scratch experiments demonstrated a significant effect of CDK1
knockdown on reducing glioma cell motility. (J) Statistical results
of the U251MG cell line migration assay (NC vs. CDK1-si, 24 h
P=0.1445; 48 h P=0.0027; 72 h P=0.0001; n=3). (K and L) Analysis of
apoptosis by flow cytometry using Annexin V-FITC and PI staining in
U251MG cells. (K) Representative dot plots and (L) quantitative
analysis (NC vs. CDK1-si, P<0.001; n=3). (M and N) Detection of
apoptosis by AO/EB fluorescence staining in U251MG cells. (M)
Representative fluorescence images (magnification, ×10) and the (N)
quantitative analysis (NC vs. CDK1-si, P=0.0089; n=3). (O and P)
Detection of apoptosis by AO/EB fluorescence staining in LN229
cells. (O) Representative fluorescence images (magnification, ×10)
and the (P) quantitative analysis (NC vs. CDK1-si, P=0.0143; n=3).
Images of γ-H2AX foci formed in (Q) U251MG and (R) LN229 cell lines
at 0.5, 2, 6 and 24 h after 5 Gy radiation. The statistical
analysis of γ-H2AX foci formed in (S) U251MG and (T) LN229 cell
lines (NC vs. CDK1-si; n=3). Data are presented as mean ± SD.
Statistical significance was calculated using unpaired t-test or
one-way analysis of variance followed by Sidak's multiple
comparisons test. *P<0.05, **P<0.01, ***P<0.001,
****P<0.0001. ns, not significant. CDK1, cyclin-dependent kinase
1; NC, negative control; si, small interfering (RNA); AO/EB,
acridine orange/ethidium bromide.

Figure 7

DEG analysis and pathway enrichment
analysis of the CDK1 gene based on Chinese Glioma Genome Atlas
dataset. (A) Volcano plots of DEGs. The red nodes represent the
significantly upregulated genes with log2FC>1 and P<0.05. The
green nodes represent the significantly downregulated genes with
log2FC<-1 and P<0.05. (B) Heatmap of DEGs indicating the
significantly upregulated and significantly downregulated DEGs. (C)
Enriched cell cycle signaling pathway by Gene Set Enrichment
Analysis. (D) Heatmap of pathway enrichment analysis across DEGs
based on Metascape analysis. FC, fold change; DEG, differentially
expressed gene; CDK1, cyclin-dependent kinase 1.

Figure 8

Association analysis between CDK1 and
cell cycle hub genes. (A) Intersecting genes of the cell cycle
signaling pathway-related genes from Metascape analysis and GSEA
are displayed by the Venn diagram. (B) CDK1 PPI network was
constructed and the top 8 hub proteins with a co-expression value
of >0.9 were screened out based on the STRING website and
Cytoscape visualization software. (C) STRING-based PPI analysis
among the top 8 hub proteins (top). Matrix correlation analysis and
visualization of CDK1 and hub genes (bottom). Scatter plot analysis
of the correlation between CDK1 and hub genes expression: (D) PCNA,
(E) MCM2, (F) MCM4, (G) MCM3, (H) MCM6, (I) PLK1, (J) TTK and (K)
MAD2L1. (L) Univariate regression analysis of the top 8 Hub genes.
Statistical significance was calculated using Pearson's correlation
coefficient. CDK1, cyclin-dependent kinase 1; GSEA, Gene Set
Enrichment Analysis; PPI, protein-protein interaction; PCNA,
proliferating cell nuclear antigen; MCM2/3/4/6, minichromosome
maintenance complex component 2/3/4/6; PLK1, polo-like kinase 1;
MAD2L1, mitotic arrest deficient 2 like 1.

Figure 9

Expression of the top 8 hub genes and
prognostic analysis. Expression level of the hub genes in glioma
and normal tissues based on the TCGA dataset alongside survival
analysis curves for (A) PCNA, (B) MCM2, (C) MCM3, (D) MCM4, (E)
MCM6, (F) PLK1, (G) TTK and (H) MAD2L1. (I-P) Quantitative PCR
results following CDK1 knock down (n=3) of (I) CDK1, (J) PCNA, (K)
MCM2, (L) MCM3, (M) MCM4, (N) MCM6, (O) PLK1 and (P) MAD2L1. Data
are presented as mean ± SD. Statistical significance was calculated
using log-rank test or Student's t-test. *P<0.05, **P<0.01,
***P<0.001. PCNA, proliferating cell nuclear antigen;
MCM2/3/4/6, minichromosome maintenance complex component 2/3/4/6;
PLK1, polo-like kinase 1; MAD2L1, mitotic arrest deficient 2 like
1; GBM, glioblastoma; LGG, low-grade glioma; NC, negative control;
si, small interfering (RNA); CDK1, cyclin-dependent kinase 1.
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Copy and paste a formatted citation
Spandidos Publications style
Wang Y, Zhou H, Han X, Zhang D, Hou L, Li H, Fan T, Li S and Xue X: <p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>. Oncol Rep 55: 41, 2026.
APA
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H. ... Xue, X. (2026). <p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>. Oncology Reports, 55, 41. https://doi.org/10.3892/or.2026.9046
MLA
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H., Fan, T., Li, S., Xue, X."<p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>". Oncology Reports 55.3 (2026): 41.
Chicago
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H., Fan, T., Li, S., Xue, X."<p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>". Oncology Reports 55, no. 3 (2026): 41. https://doi.org/10.3892/or.2026.9046
Copy and paste a formatted citation
x
Spandidos Publications style
Wang Y, Zhou H, Han X, Zhang D, Hou L, Li H, Fan T, Li S and Xue X: <p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>. Oncol Rep 55: 41, 2026.
APA
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H. ... Xue, X. (2026). <p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>. Oncology Reports, 55, 41. https://doi.org/10.3892/or.2026.9046
MLA
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H., Fan, T., Li, S., Xue, X."<p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>". Oncology Reports 55.3 (2026): 41.
Chicago
Wang, Y., Zhou, H., Han, X., Zhang, D., Hou, L., Li, H., Fan, T., Li, S., Xue, X."<p>CDK1 depletion suppresses glioma malignancy through cell cycle pathway regulation: Mechanistic insights from functional and molecular profiling</p>". Oncology Reports 55, no. 3 (2026): 41. https://doi.org/10.3892/or.2026.9046
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