Journal: Breast Cancer Research : BCR

Article Title: Neutrophil extracellular traps induced by neoadjuvant chemotherapy of breast cancer promotes vascular endothelial damage

doi: 10.1186/s13058-025-02011-y

Figure Lengend Snippet: TCb Enhances Neutrophil Extracellular Traps (NETs) Formation in Neoadjuvant Chemotherapy Patients and Animal Models. A Plasma dsDNA concentrations in normal control breast cancer patients, and those undergoing EC-T, EC-T-HP and TCb neoadjuvant chemotherapy. Plasma dsDNA levels were significantly elevated in TCb-treated patients compared to controls. NC (n = 50), EC-T (n = 50), EC-T-HP (n = 19), TCb (n = 45), B plasma Myeloperoxidas (MPO) and histone H3 concentrations in patients showed significant increases in TCb-treated patients compared to controls, C , D immunofluorescence staining showed more pronounced co-expression of MPO and histone H3 in neutrophils from TCb-treated patients than controls, E plasma dsDNA concentrations in mice from TCb chemotherapy models and controls. Plasma dsDNA was significantly elevated in TCb model mice compared to controls (n = 6), F plasma MPO and histone H3 concentrations in mouse models showed significant increases in TCb models compared to controls. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Neutrophils isolated and purified from patients undergoing neoadjuvant chemotherapy for breast cancer were stained with anti-myeloperoxidase antibody (Proteintech, USA, Catalog No. 22225–1-AP, diluted 1:50) and anti-histone H3 antibody (Proteintech, Catalog No. 17168–1-AP, diluted 1:1200) [ – ].

Techniques: Clinical Proteomics, Control, Immunofluorescence, Staining, Expressing

Journal: Breast Cancer Research : BCR

Article Title: Neutrophil extracellular traps induced by neoadjuvant chemotherapy of breast cancer promotes vascular endothelial damage

doi: 10.1186/s13058-025-02011-y

Figure Lengend Snippet: Induction of NETs Following TCb Stimulation, Demonstrating NETs-Mediated Damage in Human Umbilical Vein Endothelial Cells (HUVECs). A Schematic representation of the experimental setup, illustrating the in vitro application of the TCb chemotherapeutic agent to induce NETs formation in neutrophils and generate conditioned media containing NETs for treating HUVECs, B neutrophils extracted from healthy volunteers were treated with TCb for 4, 8, and 24 h in vitro. An increase in NETs dsDNA concentration in the cell culture supernatant was observed starting 8 h post-treatment, C After 8 h of TCb treatment, significant elevations in MPO and histone H3 concentrations were observed in the cell culture supernatant of the TCb-treated group compared to the untreated control, D , E immunofluorescence staining revealed a more pronounced co-expression of MPO and histone H3 in TCb-treated human neutrophils compared to untreated controls, F cell viability of HUVECs treated with TCb-induced NETs was assessed using the CCK-8 assay, revealing a significant decrease compared to untreated controls, G , H western blot analysis revealed significant downregulation of VE-cadherin, CD31, and Syndecan-4, and upregulation of Bax in HUVECs treated with TCb-induced NETs, indicating endothelial cell injury and apoptosis. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Neutrophils isolated and purified from patients undergoing neoadjuvant chemotherapy for breast cancer were stained with anti-myeloperoxidase antibody (Proteintech, USA, Catalog No. 22225–1-AP, diluted 1:50) and anti-histone H3 antibody (Proteintech, Catalog No. 17168–1-AP, diluted 1:1200) [ – ].

Techniques: In Vitro, Concentration Assay, Cell Culture, Control, Immunofluorescence, Staining, Expressing, CCK-8 Assay, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Neutrophil extracellular traps induced by neoadjuvant chemotherapy of breast cancer promotes vascular endothelial damage

doi: 10.1186/s13058-025-02011-y

Figure Lengend Snippet: Reversal of Vascular Endothelial Injury Markers Induced by NETs Formation in TCb Mouse Models. A Schematic representation of the mouse models used for TCb chemotherapy and CI-amidine intervention, B administration of CI-amidine to TCb mouse models reversed the elevated plasma dsDNA concentrations observed in TCb-treated mice (n = 6), C plasma levels of MPO and histone H3 in mouse models showed that CI-amidine mitigated increases induced by TCb chemotherapy, D plasma concentrations of vWF and Syndecan-4, significantly upregulated in TCb-treated mice compared to controls, were reversed by CI-amidine intervention. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Neutrophils isolated and purified from patients undergoing neoadjuvant chemotherapy for breast cancer were stained with anti-myeloperoxidase antibody (Proteintech, USA, Catalog No. 22225–1-AP, diluted 1:50) and anti-histone H3 antibody (Proteintech, Catalog No. 17168–1-AP, diluted 1:1200) [ – ].

Techniques: Clinical Proteomics

Journal: Breast Cancer Research : BCR

Article Title: Neutrophil extracellular traps induced by neoadjuvant chemotherapy of breast cancer promotes vascular endothelial damage

doi: 10.1186/s13058-025-02011-y

Figure Lengend Snippet: Slc11a1 Knockdown Reduces ROS, Ferrous Ion Contents and NETs Production in Human Neutrophils In Vitro. A Human neutrophils were treated with the TCb chemotherapeutic agent or transfected with Slc11a1 small interfering RNA (siRNA). Slc11a1 mRNA levels were measured by q-PCR. TCb treatment significantly increased Slc11a1 mRNA expression, which was reversed upon knockdown of Slc11a1 , B , C western blot analysis showed that TCb treatment significantly elevated NRAMP1 protein levels in human neutrophils, which was reversed by Slc11a1 knockdown, D – F intracellular ferrous ion levels, measured using the FerroOrange probe in human neutrophils, showed a significant increase in TCb-treated groups, attenuated by Slc11a1 knockdown, G , H ROS levels in TCb-treated human neutrophils demonstrated a marked elevation compared to controls, which was reversed by Slc11a1 knockdown, I quantification of dsDNA in cell culture supernatants showed a significant increase in TCb-treated human neutrophils, which was significantly reversed by Slc11a1 knockdown, J MPO and histone H3 concentrations in the culture supernatants of TCb-treated human neutrophils were elevated but reversed by Slc11a1 knockdown. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Neutrophils isolated and purified from patients undergoing neoadjuvant chemotherapy for breast cancer were stained with anti-myeloperoxidase antibody (Proteintech, USA, Catalog No. 22225–1-AP, diluted 1:50) and anti-histone H3 antibody (Proteintech, Catalog No. 17168–1-AP, diluted 1:1200) [ – ].

Techniques: Knockdown, In Vitro, Transfection, Small Interfering RNA, Expressing, Western Blot, Cell Culture

Journal: Breast Cancer Research : BCR

Article Title: Neutrophil extracellular traps induced by neoadjuvant chemotherapy of breast cancer promotes vascular endothelial damage

doi: 10.1186/s13058-025-02011-y

Figure Lengend Snippet: Slc11a1 Knockdown in Mouse Models Leads to Reduced ROS, Ferrous Ions and NETs in Neutrophils. A Schematic representation of generating Slc11a1 knockdown mice using AAV constructs and constructing a TCb chemotherapy mouse model, B q-PCR analysis of Slc11a1 mRNA expression in neutrophils from mouse models. Significant increases in Slc11a1 mRNA expression in neutrophils from TCb-treated mice were reversed in Slc11a1 -knockdown TCb-treated mice, C – E detection of ferrous ion levels in neutrophils from mouse models. Elevated ferrous ion content in neutrophils from TCb-treated mice was notably reversed in Slc11a1 -knockdown mice, F , G measurement of ROS levels in neutrophils from mouse models. Elevated ROS levels in neutrophils from TCb-treated mice were significantly mitigated in Slc11a1 -knockdown mice, H quantification of dsDNA in plasma from mouse models. Increased plasma dsDNA concentrations in TCb-treated mice were significantly attenuated in Slc11a1 -knockdown mice, I analysis of MPO and histone H3 levels in plasma from mouse models. Elevated concentrations of MPO and histone H3 in plasma from TCb-treated mice were significantly reversed in Slc11a1 -knockdown mice, J assessment of vWF and Syndecan-4 levels in plasma from mouse models. Increased plasma concentrations of vWF and Syndecn-4 in TCb-treated mice were significantly diminished in Slc11a1 -knockdown mice. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Neutrophils isolated and purified from patients undergoing neoadjuvant chemotherapy for breast cancer were stained with anti-myeloperoxidase antibody (Proteintech, USA, Catalog No. 22225–1-AP, diluted 1:50) and anti-histone H3 antibody (Proteintech, Catalog No. 17168–1-AP, diluted 1:1200) [ – ].

Techniques: Knockdown, Construct, Expressing, Clinical Proteomics