Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line
<b>Background:</b> The clinical relevance of circulating cell-free DNA (cfDNA) in oncology has gained significant attention, with its potential as a biomarker for cancer diagnosis and monitoring. However, its precise role in cancer biology and progression remains unclear. cfDNA in cancer...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-03-01
|
| Series: | Metabolites |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-1989/15/4/227 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849714198251044864 |
|---|---|
| author | Isabel Lemos Catarina Freitas-Dias Ana Hipólito José Ramalho Fabrizio Carteni Luís G. Gonçalves Stefano Mazzoleni Jacinta Serpa |
| author_facet | Isabel Lemos Catarina Freitas-Dias Ana Hipólito José Ramalho Fabrizio Carteni Luís G. Gonçalves Stefano Mazzoleni Jacinta Serpa |
| author_sort | Isabel Lemos |
| collection | DOAJ |
| description | <b>Background:</b> The clinical relevance of circulating cell-free DNA (cfDNA) in oncology has gained significant attention, with its potential as a biomarker for cancer diagnosis and monitoring. However, its precise role in cancer biology and progression remains unclear. cfDNA in cancer patients’ blood has been shown to activate signaling pathways, such as those mediated by toll-like receptors (TLRs), suggesting its involvement in cancer cell adaptation to the tumor microenvironment. <b>Methods:</b> This impact of cfDNA released from MDA-MB-231, a triple-negative breast cancer (TNBC) cell line was assessed, focusing on glucose availability and culture duration. The impact of cfDNA on the proliferation of MDA-MB-231 cells was investigated using proliferation curves, while cellular migration was evaluated through wound healing assays. The metabolic alterations induced by distinct cfDNA variants in MDA-MB-231 cells were investigated through nuclear magnetic resonance (NMR) spectroscopy, and their effect on cisplatin resistance was evaluated using flow cytometry. Furthermore, the expression levels of DNA-sensitive Toll-like receptor 9 (TLR9) were quantified via immunofluorescence, alongside its colocalization with lysosome-associated membrane protein 1 (LAMP1). <b>Results:</b> This study indicates that cfDNA facilitates metabolic adaptation, particularly under metabolic stress, by modulating glucose and glutamine consumption, key pathways in tumor cell metabolism. Exposure to cfDNA induced distinct metabolic shifts, favoring energy production through oxidative phosphorylation. The anti-cancer activity of cfDNA isolated from conditioned media of cells cultured under stressful conditions is influenced by the culture duration, emphasizing the importance of adaptation and se-lection in releasing cfDNA that can drive pro-tumoral processes. Additionally, cfDNA exposure influenced cell proliferation, quiescence, and migration, processes linked to metastasis and treatment resistance. These findings underscore cfDNA as a key mediator of metabolic reprogramming and adaptive responses in cancer cells, contributing to tumor progression and therapy resistance. Furthermore, the activation of TLR9 signaling suggests a mechanistic basis for cfDNA-induced phenotypic changes. <b>Conclusions</b>: Overall, cfDNA serves as a crucial signaling molecule in the tumor microenvironment, orchestrating adaptive processes that enhance cancer cell survival and progression. |
| format | Article |
| id | doaj-art-eef6aba1581545d89d839e58c70b87af |
| institution | DOAJ |
| issn | 2218-1989 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj-art-eef6aba1581545d89d839e58c70b87af2025-08-20T03:13:45ZengMDPI AGMetabolites2218-19892025-03-0115422710.3390/metabo15040227Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell LineIsabel Lemos0Catarina Freitas-Dias1Ana Hipólito2José Ramalho3Fabrizio Carteni4Luís G. Gonçalves5Stefano Mazzoleni6Jacinta Serpa7iNOVA4Health, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisbon, PortugaliNOVA4Health, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisbon, PortugaliNOVA4Health, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisbon, PortugaliNOVA4Health, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisbon, PortugalLab Applied Ecology and System Dynamics, Dipartimento di Agraria, Università di Napoli “Federico II”, Portici, 80055 Naples, ItalyInstituto de Tecnologia Química e Biológica António Xavier (ITQB NOVA), Avenida da República (EAN), 2780-157 Oeiras, PortugalLab Applied Ecology and System Dynamics, Dipartimento di Agraria, Università di Napoli “Federico II”, Portici, 80055 Naples, ItalyiNOVA4Health, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria, 130, 1169-056 Lisbon, Portugal<b>Background:</b> The clinical relevance of circulating cell-free DNA (cfDNA) in oncology has gained significant attention, with its potential as a biomarker for cancer diagnosis and monitoring. However, its precise role in cancer biology and progression remains unclear. cfDNA in cancer patients’ blood has been shown to activate signaling pathways, such as those mediated by toll-like receptors (TLRs), suggesting its involvement in cancer cell adaptation to the tumor microenvironment. <b>Methods:</b> This impact of cfDNA released from MDA-MB-231, a triple-negative breast cancer (TNBC) cell line was assessed, focusing on glucose availability and culture duration. The impact of cfDNA on the proliferation of MDA-MB-231 cells was investigated using proliferation curves, while cellular migration was evaluated through wound healing assays. The metabolic alterations induced by distinct cfDNA variants in MDA-MB-231 cells were investigated through nuclear magnetic resonance (NMR) spectroscopy, and their effect on cisplatin resistance was evaluated using flow cytometry. Furthermore, the expression levels of DNA-sensitive Toll-like receptor 9 (TLR9) were quantified via immunofluorescence, alongside its colocalization with lysosome-associated membrane protein 1 (LAMP1). <b>Results:</b> This study indicates that cfDNA facilitates metabolic adaptation, particularly under metabolic stress, by modulating glucose and glutamine consumption, key pathways in tumor cell metabolism. Exposure to cfDNA induced distinct metabolic shifts, favoring energy production through oxidative phosphorylation. The anti-cancer activity of cfDNA isolated from conditioned media of cells cultured under stressful conditions is influenced by the culture duration, emphasizing the importance of adaptation and se-lection in releasing cfDNA that can drive pro-tumoral processes. Additionally, cfDNA exposure influenced cell proliferation, quiescence, and migration, processes linked to metastasis and treatment resistance. These findings underscore cfDNA as a key mediator of metabolic reprogramming and adaptive responses in cancer cells, contributing to tumor progression and therapy resistance. Furthermore, the activation of TLR9 signaling suggests a mechanistic basis for cfDNA-induced phenotypic changes. <b>Conclusions</b>: Overall, cfDNA serves as a crucial signaling molecule in the tumor microenvironment, orchestrating adaptive processes that enhance cancer cell survival and progression.https://www.mdpi.com/2218-1989/15/4/227cell-free DNA (cfDNA)metabolic remodelingcell proliferationquiescencemigrationtoll-like receptor 9 (TLR9) |
| spellingShingle | Isabel Lemos Catarina Freitas-Dias Ana Hipólito José Ramalho Fabrizio Carteni Luís G. Gonçalves Stefano Mazzoleni Jacinta Serpa Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line Metabolites cell-free DNA (cfDNA) metabolic remodeling cell proliferation quiescence migration toll-like receptor 9 (TLR9) |
| title | Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line |
| title_full | Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line |
| title_fullStr | Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line |
| title_full_unstemmed | Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line |
| title_short | Cell-Free DNA (cfDNA) Regulates Metabolic Remodeling, Sustaining Proliferation, Quiescence, and Migration in MDA-MB-231, a Triple-Negative Breast Carcinoma (TNBC) Cell Line |
| title_sort | cell free dna cfdna regulates metabolic remodeling sustaining proliferation quiescence and migration in mda mb 231 a triple negative breast carcinoma tnbc cell line |
| topic | cell-free DNA (cfDNA) metabolic remodeling cell proliferation quiescence migration toll-like receptor 9 (TLR9) |
| url | https://www.mdpi.com/2218-1989/15/4/227 |
| work_keys_str_mv | AT isabellemos cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT catarinafreitasdias cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT anahipolito cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT joseramalho cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT fabriziocarteni cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT luisggoncalves cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT stefanomazzoleni cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline AT jacintaserpa cellfreednacfdnaregulatesmetabolicremodelingsustainingproliferationquiescenceandmigrationinmdamb231atriplenegativebreastcarcinomatnbccellline |