Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways

Background: Brain metastasis (BrM) is a common complication of advanced tumors with poor prognosis. Although radiotherapy remains a key treatment for BrM, it is plagued by issues such as radiation-induced brain necrosis, neurocognitive impairment, and progress post-treatment. Tumor Treating Fields (...

Full description

Saved in:
Bibliographic Details
Main Authors: Guilong Tanzhu, Haiqin Peng, Liu Chen, Gang Xiao, Jiaoyang Ning, Ling Chen, Rongrong Zhou
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Brain Stimulation
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S1935861X25002864
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849236240819290112
author Guilong Tanzhu
Haiqin Peng
Liu Chen
Gang Xiao
Jiaoyang Ning
Ling Chen
Rongrong Zhou
author_facet Guilong Tanzhu
Haiqin Peng
Liu Chen
Gang Xiao
Jiaoyang Ning
Ling Chen
Rongrong Zhou
author_sort Guilong Tanzhu
collection DOAJ
description Background: Brain metastasis (BrM) is a common complication of advanced tumors with poor prognosis. Although radiotherapy remains a key treatment for BrM, it is plagued by issues such as radiation-induced brain necrosis, neurocognitive impairment, and progress post-treatment. Tumor Treating Fields (TTFields) therapy employs medium frequency (100∼300 kHz) and low intensity (1∼3 v/cm) alternating electric fields to inhibit tumors. We explored the effects and mechanisms of TTFields combined with ionizing radiation (IR) on the malignant phenotype of lung cancer brain metastasis (LCBM) cells, with the aim of advancing the clinical adoption of TTFields. Methods: LCBM cells H1915, PC9-Brm and primary cells were used. The experiments included four groups: Control, 6 Gy, TTFields, and 6 Gy + TTFields group. Cell viability and the number of EDU or Ki67-positive cells, ability of migration and colony formation were assessed; Cell morphology was observed by H&E staining; Bulk transcriptome revealed the potential mechanisms, with the protein levels of differentially expressed genes (DEGs) verified through immunofluorescence assay. DNA damage repair pathways were validated by Western Blot. Additionally, ROS levels, and HO-1 expression were evaluated. Results: The combination of 150 kHz, 2∼2.5 v/cm TTFields and 6 Gy IR effectively suppressed the viability and the number of EDU-positive and Ki67-positive LCBM cells. Following TTFields and IR, the tumor cells exhibited altered morphology and reduced clonogenic and migratory capacities. RNA-seq revealed associations between TTFields combined with IR and various biological processes and mechanisms, including redox reactions (primarily related to mitochondria), DNA replication, transition metal ion transmembrane transport, and heme metabolism. Furthermore, TTFields combined with IR enhanced γH2AX and 53BP1 levels. Increased ROS and HO-1 expression were observed post-treatment. Similarly, LCBM primary cells exhibited decreased Ki67-positive cells along with increased γH2AX and 53BP1 foci following TTFields and IR. TTFields combined with IR exhibited significant suppression on homologous recombination (HR) markers (p-ATM, RAD51), non-homologous end joining (NHEJ) components (DNA-PKcs, KU70, KU80), and microhomology-mediated end joining (MMEJ) effectors (PARP1, p95-NBS1) versus RT alone. Conclusions: TTFields combined with IR effectively inhibits LCBM cells. Mechanistically, this combined treatment enhanced DNA damage, suppressed DNA damage repair, and elevates ROS levels.
format Article
id doaj-art-cdd76a1d937d4df39a3b386f4cce30c4
institution Kabale University
issn 1935-861X
language English
publishDate 2025-09-01
publisher Elsevier
record_format Article
series Brain Stimulation
spelling doaj-art-cdd76a1d937d4df39a3b386f4cce30c42025-08-20T04:02:23ZengElsevierBrain Stimulation1935-861X2025-09-011851426144010.1016/j.brs.2025.07.014Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathwaysGuilong Tanzhu0Haiqin Peng1Liu Chen2Gang Xiao3Jiaoyang Ning4Ling Chen5Rongrong Zhou6Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaDepartment of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaDepartment of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaDepartment of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaDepartment of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaDepartment of Neurosurgery, Chinese People's Liberation Army of China (PLA) General Hospital, Medical School of Chinese PLA, Institute of Neurosurgery of Chinese PLA, 100853, Beijing, China; Corresponding author.Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, China; Xiangya Lung Cancer Center, Xiangya Hospital, Central South University, Changsha, 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, Hunan Province, 410008, China; Corresponding author.Department of Oncology, Xiangya Hospital, Central South University, Changsha, 410008, ChinaBackground: Brain metastasis (BrM) is a common complication of advanced tumors with poor prognosis. Although radiotherapy remains a key treatment for BrM, it is plagued by issues such as radiation-induced brain necrosis, neurocognitive impairment, and progress post-treatment. Tumor Treating Fields (TTFields) therapy employs medium frequency (100∼300 kHz) and low intensity (1∼3 v/cm) alternating electric fields to inhibit tumors. We explored the effects and mechanisms of TTFields combined with ionizing radiation (IR) on the malignant phenotype of lung cancer brain metastasis (LCBM) cells, with the aim of advancing the clinical adoption of TTFields. Methods: LCBM cells H1915, PC9-Brm and primary cells were used. The experiments included four groups: Control, 6 Gy, TTFields, and 6 Gy + TTFields group. Cell viability and the number of EDU or Ki67-positive cells, ability of migration and colony formation were assessed; Cell morphology was observed by H&E staining; Bulk transcriptome revealed the potential mechanisms, with the protein levels of differentially expressed genes (DEGs) verified through immunofluorescence assay. DNA damage repair pathways were validated by Western Blot. Additionally, ROS levels, and HO-1 expression were evaluated. Results: The combination of 150 kHz, 2∼2.5 v/cm TTFields and 6 Gy IR effectively suppressed the viability and the number of EDU-positive and Ki67-positive LCBM cells. Following TTFields and IR, the tumor cells exhibited altered morphology and reduced clonogenic and migratory capacities. RNA-seq revealed associations between TTFields combined with IR and various biological processes and mechanisms, including redox reactions (primarily related to mitochondria), DNA replication, transition metal ion transmembrane transport, and heme metabolism. Furthermore, TTFields combined with IR enhanced γH2AX and 53BP1 levels. Increased ROS and HO-1 expression were observed post-treatment. Similarly, LCBM primary cells exhibited decreased Ki67-positive cells along with increased γH2AX and 53BP1 foci following TTFields and IR. TTFields combined with IR exhibited significant suppression on homologous recombination (HR) markers (p-ATM, RAD51), non-homologous end joining (NHEJ) components (DNA-PKcs, KU70, KU80), and microhomology-mediated end joining (MMEJ) effectors (PARP1, p95-NBS1) versus RT alone. Conclusions: TTFields combined with IR effectively inhibits LCBM cells. Mechanistically, this combined treatment enhanced DNA damage, suppressed DNA damage repair, and elevates ROS levels.http://www.sciencedirect.com/science/article/pii/S1935861X25002864Lung cancer brain metastasisTTFieldsRadiotherapyDNA damageMolecular mechanisms
spellingShingle Guilong Tanzhu
Haiqin Peng
Liu Chen
Gang Xiao
Jiaoyang Ning
Ling Chen
Rongrong Zhou
Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
Brain Stimulation
Lung cancer brain metastasis
TTFields
Radiotherapy
DNA damage
Molecular mechanisms
title Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
title_full Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
title_fullStr Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
title_full_unstemmed Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
title_short Tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing DNA damage repair pathways
title_sort tumor treating fields combined with ionizing radiation inhibit the malignant phenotype of lung cancer brain metastasis cells by suppressing dna damage repair pathways
topic Lung cancer brain metastasis
TTFields
Radiotherapy
DNA damage
Molecular mechanisms
url http://www.sciencedirect.com/science/article/pii/S1935861X25002864
work_keys_str_mv AT guilongtanzhu tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT haiqinpeng tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT liuchen tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT gangxiao tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT jiaoyangning tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT lingchen tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways
AT rongrongzhou tumortreatingfieldscombinedwithionizingradiationinhibitthemalignantphenotypeoflungcancerbrainmetastasiscellsbysuppressingdnadamagerepairpathways