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Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines

  • Authors:
    • Jinju Heo
    • Myonggeun Yoon
  • View Affiliations / Copyright

    Affiliations: Department of Biomedical Engineering, Korea University, Seoul 02841, Republic of Korea
    Copyright: © Heo et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 200
    |
    Published online on: May 29, 2026
       https://doi.org/10.3892/etm.2026.13195
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Abstract

T‑cell acute lymphoblastic leukemia (T‑ALL) is a highly aggressive hematologic malignancy with limited treatment options for relapsed or refractory disease. Tumor Treating Fields (TTFields), low‑intensity alternating electric fields that disrupt mitotic spindle assembly, have been clinically determined in numerous solid tumors, yet their effects in hematologic malignancies remain incompletely understood. In the present study, the cellular responses of T‑ALL cells to TTFields were examined. Jurkat and MOLT‑4 cells were exposed to 150 kHz and 0.7 V/cm fields for up to 96 h. TTFields treatment progressively suppressed cell proliferation, induced G2/M‑phase arrest, altered cell‑cycle distribution, increased apoptotic fractions and increased late‑apoptotic fractions. TTFields exposure also increased side‑scatter intensity (indicative of increased cellular granularity or internal complexity) and MitoTracker green fluorescence in both cell lines. Intracellular ATP levels were reduced in Jurkat cells, whereas no significant change was observed in MOLT‑4 cells. In parallel, transcriptional analyses revealed downregulation of CD69 and IL‑2 and upregulation of RELA proto‑oncogene, NF‑κB subunit and CBL proto‑oncogene B. Collectively, these results demonstrate that TTFields induces cytostatic and apoptotic effects accompanied by measurable structural, bioenergetic and transcriptional alterations in T‑ALL cells, supporting further investigation of TTFields as a physical therapeutic approach for hematologic malignancies.
View Figures

Figure 1

TTFields reduce viability of T-cell
acute lymphoblastic leukemia cells in an intensity- and
time-dependent manner. (A) Normalized cell viability of Jurkat
(black) and MOLT-4 (gray) cells after exposure to 0.0-1.2 V/cm
TTFields. Jurkat exhibits minimal change at 0.4 V/cm but sharp
reductions at ≥0.8 V/cm; MOLT-4 declines across all intensities.
The right panel shows cell viability across the tested frequency
range, with no marked frequency-dependent differences observed. (B)
Time-course of live-cell counts (0-96 h) under control vs.
TTFields. Error bars represent the mean ± SD of three independent
experiments. *P<0.05, **P<0.01 and
***P<0.001. TTFields/TTF, Tumor Treating Fields.

Figure 2

Effect of TTFields on cell-cycle
distribution and apoptosis in T-ALL cell lines (A) Cell-cycle
analysis showing an increased sub-G1 fraction and a
decreased G1 phase population following TTFields
treatment, indicating increased apoptotic fractions. (B) Annexin
V/PI staining demonstrates elevated late-apoptotic cell populations
and reduced viable fractions. Data are presented as the mean ± SD
from three independent experiments. **P<0.01 and
***P<0.001. TTFields/TTF, Tumor Treating Fields;
PE-A, phycoerythrin-area; Con, control.

Figure 3

TTFields increase cellular granularity
and mitochondrial fluorescence in T-cell acute lymphoblastic
leukemia cell lines. (A) Representative FSC/SSC plots and
quantification of side-scatter mean fluorescence intensity (median
SSC-A values). FSC-A associates with cell size, while SSC-A
reflects intracellular complexity such as organelle content and
granularity. (B) MitoTracker green fluorescence intensity was
normalized to control values and was significantly elevated in both
Jurkat and MOLT-4 cells following TTFields treatment. (C)
Intracellular ATP levels measured using the CellTiter-Glo
luminescence assay and normalized to control values. Data are
presented as the mean ± SD from three independent experiments.
*P<0.05, **P<0.01 and
***P<0.001. TTFields/TTF, Tumor Treating Fields; SSC,
side scatter; FSC, forward scatter; A, area; CON, control; MFI,
mean fluorescence intensity; a.u., arbitrary units.

Figure 4

TTFields reduce CD69 expression and
alter immune-related gene transcription in T-cell acute
lymphoblastic leukemia cells. (A) Flow-cytometric analysis showed
reduced CD69+ fractions in pre-activated Jurkat and
MOLT-4 cells following 48-96 h TTFields exposure, with quantitative
summaries indicating a consistent downward trend. TTFields
treatment reduced CD69 expression in MOLT-4 cells, whereas no
significant change was observed in Jurkat cells. (B)
Gene-expression profiling after 48 h further demonstrated decreased
CD69 and IL-2 and increased RELA and CBLB transcripts. Data
represent the mean ± SD from three independent experiments.
*P<0.05, **P<0.01 and
***P<0.001. TTFields/TTF, Tumor Treating Fields;
RELA, RELA proto-oncogene, NF-κB subunit; CBLB, CBL proto-oncogene
B; Con, control.
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Copy and paste a formatted citation
Spandidos Publications style
Heo J and Yoon M: Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines. Exp Ther Med 32: 200, 2026.
APA
Heo, J., & Yoon, M. (2026). Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines. Experimental and Therapeutic Medicine, 32, 200. https://doi.org/10.3892/etm.2026.13195
MLA
Heo, J., Yoon, M."Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines". Experimental and Therapeutic Medicine 32.2 (2026): 200.
Chicago
Heo, J., Yoon, M."Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines". Experimental and Therapeutic Medicine 32, no. 2 (2026): 200. https://doi.org/10.3892/etm.2026.13195
Copy and paste a formatted citation
x
Spandidos Publications style
Heo J and Yoon M: Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines. Exp Ther Med 32: 200, 2026.
APA
Heo, J., & Yoon, M. (2026). Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines. Experimental and Therapeutic Medicine, 32, 200. https://doi.org/10.3892/etm.2026.13195
MLA
Heo, J., Yoon, M."Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines". Experimental and Therapeutic Medicine 32.2 (2026): 200.
Chicago
Heo, J., Yoon, M."Tumor Treating Fields modulate apoptotic and immune programs in T‑cell acute lymphoblastic leukemia cell lines". Experimental and Therapeutic Medicine 32, no. 2 (2026): 200. https://doi.org/10.3892/etm.2026.13195
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