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Article

A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma 

  • Authors:
    • Wenyu Liu
    • Tingting Zhang
    • Ichiyo Shibahara
    • Takuichiro Hide
    • Toshihiro Kumabe
    • Shun-Ichiro Ogura
    • Tetsuya Taga
    • Kouichi Tabu
  • View Affiliations / Copyright

    Affiliations: Department of Stem Cell Regulation, Division of Visionary Life Science, Medical Research Laboratory, Institute of Integrated Research, Institute of Science Tokyo, Tokyo 113‑8510, Japan, Department of Neurosurgery, Kitasato University School of Medicine, Sagamihara, Kanagawa 252‑0374, Japan, School of Life Science and Technology, Institute of Science Tokyo, Yokohama, Kanagawa 226‑8501, Japan
  • Article Number: 49
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    Published online on: May 14, 2026
       https://doi.org/10.3892/mco.2026.2958
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Abstract

Surgical resection remains the cornerstone of the standard therapy for glioblastoma (GBM), yet postoperative recurrence remains a major clinical challenge. Intraoperative photodynamic detection (PDD) using 5‑aminolevulinic acid (5‑ALA) improves the precision of tumor removal. However, a subset of tumor cells can evade fluorescence‑based detection, potentially contributing to residual disease and relapse. The present study analyzed the relationship between cellular migratory capacity and 5‑ALA PDD visibility using six patient‑derived GBM cell lines (KBT#12137, KBT#10135, KBT#10170, PDM19, PDM22 and PDM123). This study revealed a positive correlation, indicating that cells with higher migratory potential accumulated lower levels of 5‑ALA‑induced protoporphyrin IX and were therefore more likely to escape intraoperative detection. Transcriptomic profiling identified gene expression signatures specifically associated with this phenotype, highlighting pathways related to cytoskeletal regulation, heme metabolism, and transporter activity. These findings suggest that the migratory potential and diagnostic evasion are functionally linked through shared molecular programs, thereby providing a potential basis for identifying predictive biomarkers associated with incomplete resection and recurrence risk. Overall, this study provides insights that may contribute to improved intraoperative strategies and targeted therapeutic approaches for GBM.
View Figures

Figure 1

Correlation between PpIX accumulation
and migratory potential in patient-derived GBM cells. (A)
Representative flow cytometry histograms showing 5-ALA-induced PpIX
fluorescence in the recurrent GBM line PDM123. (B) Quantification
of PpIX-negative cell fractions in three primary (KBT series) and
three recurrent (PDM series) GBM lines. KBT#12137 showed the
highest PpIX accumulation, whereas PDM123 exhibited the lowest,
consistent with its recurrent origin. Significant differences were
observed between the primary and recurrent groups.
*P<0.05. (C) Quantitative analysis of migratory
capacity assessed by Transwell assay across the six GBM lines.
Recurrent lines displayed significantly higher motility than
primary lines. ***P<0.001. (D) Correlation between
the proportion (%) of PpIX-negative cells and the migratory index,
defined as the migration rate (%)=(lower-/upper-chamber cell
counts) x100. All data represent means ± SD from at least three
independent experiments. GBM, glioblastoma; 5-ALA, 5-aminolevulinic
acid; PpIX, protoporphyrin IX.

Figure 2

Reduced PpIX accumulation in highly
migratory GBM subpopulations. (A) Representative comparison of
5-ALA-induced PpIX fluorescence between migratory (lower-chamber)
and non-migratory (upper-chamber) cells isolated from the recurrent
GBM line PDM123. The migratory fraction contained a higher
proportion of PpIX-negative cells. (B) Quantitative summary of
PpIX-negative cell fractions in migratory and non-migratory
subpopulations across six GBM lines (three primary and three
recurrent). Except for PDM19, all lines exhibited a significantly
higher frequency of PpIX-negative cells in the migratory fraction.
*P<0.05 and **P<0.01. Data represent
means ± SD from at least three independent experiments. GBM,
glioblastoma; 5-ALA, 5-aminolevulinic acid; PpIX, protoporphyrin
IX; n.s., not significant.

Figure 3

Transcriptomic and clinical
significance of the migratory PpIX-low phenotype. (A) MA plot
illustrating differential gene expression among the four
subpopulations (non-migratory/PpIX-low, non-migratory/PpIX-high,
migratory/PpIX-low, and migratory/PpIX-high) derived from the
recurrent GBM line PDM123. Genes upregulated (>2-fold) in
migratory PpIX-low cells are shown in red, downregulated
(<0.5-fold) in blue. The top five genes by |log2FC|
are annotated. (B) Heatmap of the top 200 differentially expressed
genes ranked by |log2FC| across the four subpopulations
(Group A: non-migratory/PpIX-low, B: non-migratory/PpIX-high, C:
migratory/PpIX-low, and D: migratory/PpIX-high). Migratory/PpIX-low
(Group C) cells showed a global downregulation pattern compared
with the others. The colour scale represents z-score normalised
expression. (C) Gene set enrichment analysis (GSEA) (31) dot plot summarizing the top 15
enriched pathways ranked by NES. Up- and down-regulated pathways
are displayed in the lower and upper panels, respectively. Dot
color indicates adjusted P-value (FDR), and dot size corresponds to
gene count. Full results for the top 25 upregulated and
downregulated pathways are provided in Table SI. (D) Kaplan-Meier survival
analysis of 525 patients with GBM in The Cancer Genome Atlas cohort
stratified by the migratory PpIX-low gene signature score.
High-score patients (n=263) exhibited significantly shorter overall
survival than low-score ones (n=262) (P<0.05, log-rank test).
NES, normalized enrichment score; GBM, glioblastoma; 5-ALA,
5-aminolevulinic acid; PpIX, protoporphyrin IX.
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Spandidos Publications style
Liu W, Zhang T, Shibahara I, Hide T, Kumabe T, Ogura S, Taga T and Tabu K: A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;. Mol Clin Oncol 25: 49, 2026.
APA
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S. ... Tabu, K. (2026). A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;. Molecular and Clinical Oncology, 25, 49. https://doi.org/10.3892/mco.2026.2958
MLA
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S., Taga, T., Tabu, K."A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;". Molecular and Clinical Oncology 25.1 (2026): 49.
Chicago
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S., Taga, T., Tabu, K."A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;". Molecular and Clinical Oncology 25, no. 1 (2026): 49. https://doi.org/10.3892/mco.2026.2958
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Spandidos Publications style
Liu W, Zhang T, Shibahara I, Hide T, Kumabe T, Ogura S, Taga T and Tabu K: A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;. Mol Clin Oncol 25: 49, 2026.
APA
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S. ... Tabu, K. (2026). A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;. Molecular and Clinical Oncology, 25, 49. https://doi.org/10.3892/mco.2026.2958
MLA
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S., Taga, T., Tabu, K."A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;". Molecular and Clinical Oncology 25.1 (2026): 49.
Chicago
Liu, W., Zhang, T., Shibahara, I., Hide, T., Kumabe, T., Ogura, S., Taga, T., Tabu, K."A transcriptional network underlying migratory cellular states and reduced 5‑ALA‑based photodynamic detectability in glioblastoma&nbsp;". Molecular and Clinical Oncology 25, no. 1 (2026): 49. https://doi.org/10.3892/mco.2026.2958
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