Journal: International Journal of Molecular Medicine

Article Title: Ebastine targets HER2/HER3 signaling and cancer stem cell traits to overcome trastuzumab resistance in HER2-positive breast cancer

doi: 10.3892/ijmm.2026.5751

Figure Lengend Snippet: EBA impairs cancer stem cell-like properties. (A) BT474 and SKBR3 cells were treated with EBA for 48 h, and ALDH1 activity was assessed by flow cytometry using the Aldefluor assay. DEAB was used to define the baseline of Aldefluor-positive fluorescence. (B) BT474 cells (5x10 4 cells/ml) were plated in ultra-low attachment dishes and cultured in the presence or absence of EBA for 5 days. The number and volume of mammospheres were measured by microscopy. (C) Overall survival of patients with breast cancer stratified by the co-expression of ALDH1A1 and CD44. (D) Spearman correlation analysis of ALDH1A1 and CD44 mRNA levels in patients with HER2-positive breast cancer from The Cancer Genome Atlas cohort (n=76). Kaplan-Meier survival analyses of patients with HER2-overexpressing breast cancer stratified by (E) ALDH1A1 and (F) CD44 expression. Patients were divided into high- and low-expression groups based on the median gene expression. Statistical significance was determined using the log-rank test. (G) JIMT-1 cells were treated with EBA (3 μ M) for 48 h and the CD44 high /CD24 low cell populations were identified by flow cytometry. (H) JIMT-1 cells (1.5x10 4 cells/ml) were cultured under serum-free suspension conditions in the presence of EBA (3 μ M) for 8 days. Mammosphere number and volumes were quantified. ** P<0.01 and **** P<0.0001 vs. vehicle-treated control (0 μ M EBA). EBA, ebastine; ALDH, aldehyde dehydrogenase; DEAB, diethylaminobenzaldehyde; CTL, control; ISO, isotype.

Article Snippet: The human breast cancer cell lines SKBR3, BT474, MDA-MB-453 (American Type Culture Collection) and JIMT-1 (Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH) were cultured in DMEM, MEM or RPMI-1640 (all Sigma-Aldrich; Merck KGaA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay, Flow Cytometry, Fluorescence, Cell Culture, Microscopy, Expressing, Gene Expression, Suspension, Control

Journal: Aging Cell

Article Title: Decreased Glucose Metabolism and Declined Chaperones Are Unique Features Required for the Survival of Senescent Fibroblasts and Pyruvate Dehydrogenase Is a Potent Senolytic Target

doi: 10.1111/acel.70434

Figure Lengend Snippet: Inhibiting PDH, GLS1 and Hsp90 by the combination of CPI‐613+BPTES+17‐AAG gave rise to enhanced senolysis on senescent fibroblasts as well as the therapy‐induced senescent tumor cells. (A, B) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating (A) and senescent (B) BJ cells. For the dose of each compound in use, see the results 2.7 section for more details. ** p < 0.01 by Student's t ‐test. (C, D) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating and Dox‐induced senescent lung adenocarcinoma A549 cells. *** p < 0.001 by Student's t ‐test. (E, F) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating and Dox‐induced senescent cervical carcinoma HeLa cells. *** p < 0.001 by Student's t ‐test. (G) The morphological changes of senescent BJ induced by IR, senescent A549 and HeLa cells induced by Dox at the indicated time of CPI‐613+BPTES+17‐AAG treatment under the light microscopy. The senescent cells without treatment were stained with SA‐β‐gal. Cells were imaged at magnification 200×. (H) The schematic summarization of our findings. The activities of TCA cycle and chaperones are reduced in DNA damage induced senescent cells. Co‐inhibiting Hsp90 and TCA cycle with 17‐AAG+CPI‐613+BPTES combination leads to enhanced selective elimination of senescent cells, hinting TCA cycle and glutaminolysis are novel and potent targets for senolysis.

Article Snippet: Human fibroblasts BJ (RRID: CVCL_3653; ATCC Cat#CRL‐2522), IMR‐90 (RRID: CVCL_0347; ATCC Cat#CCL‐186), WI‐38 (RRID: CVCL_0579; ATCC Cat#CCL‐75) and human cancer cell lines HeLa (RRID: CVCL_0030; ATCC Cat#CRM‐CCL‐2), A549 (RRID: CVCL_0023; ATCC Cat#CRM‐CCL‐185), and U2OS (RRID: CVCL_0042; ATCC Cat#HTB‐96) were purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin at 37°C under 5% CO 2 .

Techniques: Light Microscopy, Staining

Journal: Aging Cell

Article Title: Decreased Glucose Metabolism and Declined Chaperones Are Unique Features Required for the Survival of Senescent Fibroblasts and Pyruvate Dehydrogenase Is a Potent Senolytic Target

doi: 10.1111/acel.70434

Figure Lengend Snippet: Distinct proteomic and transcriptional signatures of metabolic enzymes and chaperones between senescent fibroblasts and the therapy‐induced senescent tumor cells. (A) The abundance of glycolysis‐related enzymes PFKP, ALDOA, PKM2, TCA cycle‐related PDHA, and glutaminolysis‐related GLS1 was decreased significantly in senescent BJ and IMR‐90 cells compared to their proliferating counterparts. (B) The protein levels of chaperones TCP1, Hsp70, and Hsp90 were decreased significantly in senescent BJ and IMR‐90 cells. (C) The abundance of those glycolysis‐related enzymes remained unchanged or even elevated in Dox‐induced senescent A549, HeLa, and U2OS tumor cells compared to their proliferating counterparts. (D) The abundance of these chaperone proteins in Dox‐induced senescent A549, HeLa, and U2OS tumor cells remained nearly unchanged. The relative abundance of each protein was quantified by signal density scanning on Western blots and normalized to the signal of β‐Actin or β‐tubulin. * p < 0.05, ** p < 0.01 tested by Student's t ‐test.

Article Snippet: Human fibroblasts BJ (RRID: CVCL_3653; ATCC Cat#CRL‐2522), IMR‐90 (RRID: CVCL_0347; ATCC Cat#CCL‐186), WI‐38 (RRID: CVCL_0579; ATCC Cat#CCL‐75) and human cancer cell lines HeLa (RRID: CVCL_0030; ATCC Cat#CRM‐CCL‐2), A549 (RRID: CVCL_0023; ATCC Cat#CRM‐CCL‐185), and U2OS (RRID: CVCL_0042; ATCC Cat#HTB‐96) were purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin at 37°C under 5% CO 2 .

Techniques: Western Blot

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces tumor cell death via frequency- and mode-dependent mechanical disruption (A) TEM images of MagGO synthesized under MF. The dashed white line represents the contour edge of GO. Scale bars, 200 nm. (B) M-H curve of GO, MNP, and MagGO. (C) AFM image of MagGO and the height of GO in MagGO. Scale bars, 500 nm. (D, F, and H) Cell viability of U87 (D), MDA-MB-231 (F), and A549 (H) cells treated with MNP and MagGO under RMF and 3D MF (RMF combined with OMF stimulation) at 5 Hz. The applied field strength is 75 mT. The data were presented as the mean ± SD. (E, G, and I) Cell viability of U87 (E), MDA-MB-231 (G), and A549 (I) cells treated with MNP and MagGO under 3D MF of different frequencies. The applied field strength is 75 mT. The data were presented as the mean ± SD.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Disruption, Synthesized

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO induces lysosomal disruption (A) CLSM images of MNPs and MagGO in U87 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (B and C) Intensity profiles (white dashed line) of signals from lysosome and MNPs/MagGO fluorescent channels in (A). (D) CLSM images of MNPs and MagGO in MDA-MB-231 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (E and F) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (D). (G) CLSM images of MNPs and MagGO in A549 cells. Lysosomes were stained with LysoTracker red (red), and the MNPs and MagGO were labeled with FITC (green). Scale bars, 15 μm. (H and I) Intensity profiles (white dashed line) of signals from lysosomes and MNPs/MagGO fluorescent channels in (G). (J) CLSM images of U87 cells transfected with the EGFP-Gal3 plasmid after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 15 μm. (K) Counts of Gal3 puncta per U87 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (L) Counts of Gal3 puncta per MDA-MB-231 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (M) Counts of Gal3 puncta per A549 cell ( n = 10). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (N) Bio-TEM of lysosomal membrane morphology after mechanoporation for MagGO and MagGO+3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 1 μm. The dark blue arrow indicates the site of LMP, while the length of the blue arrow represents the size of the lysosomal membrane “wound”.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Disruption, Staining, Labeling, Transfection, Plasmid Preparation, Membrane

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: MagGO primarily induces pyroptosis as the mode of cell death (A–C) CLSM images of CTSB release of U87 (A), MDA-MB-231 (B), and A549 (C) cells after MagGO treatment under 3D MF. The applied field strength is 75 mT. The duration of magnetic field application is 30 min. Scale bars, 20 μm. (D–F) The cell viabilities of U87 (D), MDA-MB-231 (E), and A549 (F) cells treated with MagGO were assessed after pre-treatment with z-VAD-FMK (10 μM), Necrostatin-1 (10 μM), 3-Methyladenine (10 μM), Ferrostatin-1 (2 μM), and MCC950 (10 nM) for 4 h, followed by exposure to 3D MF at 5 Hz for 30 min. The applied field strength was 75 mT. The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (G and H) Quantification of IL-1β (G) and IL-18 (H) release from U87 cells for control, MagGO, and MagGO+3D MF ( n = 3). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post-hoc test. (I and J) Western blot analysis of Casp-1 (I) and GSDMD (J) in U87 cells for control, MagGO, and MagGO+3D MF.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Control, Western Blot

Journal: iScience

Article Title: An atom-edged magnetic nanomotor for cancer mechanotherapy

doi: 10.1016/j.isci.2026.114994

Figure Lengend Snippet: Broad antitumor activity of MagGO across multiple tumor models (A) Schematic illustrations of in vivo anticancer therapy for MDA-MB-231 and A549 tumors. The applied field strength is 75 mT. The applied field frequency is 5 Hz. The duration of magnetic field application is 30 min. (B) The MDA-MB-231 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (C) The MDA-MB-231 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (D) The MDA-MB-231 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (E) The MDA-MB-231 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days. (F) The A549 tumor volume comparison of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF for 14 days ( n = 5). (G) The A549 tumor images of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). (H) The A549 tumor weight of control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF ( n = 5). The data were presented as the mean ± SD. Data were analyzed by one-way ANOVA with Tukey’s post hoc test. (I) The A549 tumor volume of each mouse in the control, MNP, MagGO, MNP+3D MF, and MagGO+3D MF groups for 14 days.

Article Snippet: Glioblastoma U87 cells, human breast cancer cell line MDA-MB-231 cells, and human lung adenocarcinoma cell line A549 cells (American Type Culture Collection, Manassas, VA, USA) were cultured at 37°C under a humidified atmosphere containing 5% CO 2 with culture medium DMEM (Hyclone) containing of 1% penicillin and streptomycin (Hyclone) and 10% fetal bovine serum (FBS, Gibco).

Techniques: Activity Assay, In Vivo, Comparison, Control