human breast cancer cell lines skbr3  (ATCC)

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: Journal of Genetic Engineering & Biotechnology

Article Title: Potential role of Adansonia digitata nanoparticles on colorectal cancer induced by colorectal cancer cells (SW620) in nude mice

doi: 10.1016/j.jgeb.2026.100659

Figure Lengend Snippet: Oxidative stress markers in nude mice colon treated with SW620, AD, ADNPs1, and ADNPs2. A significant change at p < 0.05 in MDA and a non-significant change at p > 0.05 in SOD and catalase activity. Values presented as mean ± S.E., a, b different superscripts within columns are significantly different ( P < 0.05).

Article Snippet: The SW620 human colorectal cancer cell line was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Activity Assay

Journal: Journal of Genetic Engineering & Biotechnology

Article Title: Potential role of Adansonia digitata nanoparticles on colorectal cancer induced by colorectal cancer cells (SW620) in nude mice

doi: 10.1016/j.jgeb.2026.100659

Figure Lengend Snippet: Anti-apoptotic/pro-apoptotic protein level markers in nude mice colon treated with SW620, AD, ADNPs1, and ADNPs2. A significant change at p < 0.05 compared to the SW620 group in cytochrome c and a non-significant change at p > 0.05 in caspase3, BAX, and BCL-2. Values presented as mean ± S.E., a, b different superscripts within columns are significantly different ( P < 0.05).

Article Snippet: The SW620 human colorectal cancer cell line was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques:

Journal: Journal of Genetic Engineering & Biotechnology

Article Title: Potential role of Adansonia digitata nanoparticles on colorectal cancer induced by colorectal cancer cells (SW620) in nude mice

doi: 10.1016/j.jgeb.2026.100659

Figure Lengend Snippet: TGF-β, TNF-α, INOS, and IL-1β mRNA expression in nude mice colon treated with SW620, AD, ADNPs1, and ADNPs2. A significant change at p < 0.05 compared to the SW620 group in TGF-β, and a non-significant change at p > 0.05 in TNF-α, IL-1β, and INOS. Values presented as mean ± S.E., a, b, c different superscripts within columns are significantly different ( P < 0.05).

Article Snippet: The SW620 human colorectal cancer cell line was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).

Techniques: Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Radiotherapy enhances M1 macrophage immunogenic activity through IFNs induction and stimulation in TP53-wild type tumors

doi: 10.1007/s00262-026-04300-7

Figure Lengend Snippet: IFNs mediate immunotherapy-associated gene expression in tumors and activate immune cells in healthy PBMCs. A qPCR was used to detect the PD-L1 expression in A549 (5 × 10 5 in a 6-well plate) treated with IFNα and IFNγ (20 ng/mL for each, incubated for 2 h). B The expression of the top 8 up-regulated genes from RNAseq analysis (Table S2), including ICAM1, BATF, IRF1, SOCS1, HAPLN3, TAP1, PSMB9, and MAFF, were validated by qPCR analysis in A549 treated with IFNα and IFNγ (20 ng/mL for each, incubated for 2 h). C The healthy PBMCs (1 × 10 6 in a 6-well plate) treated with IFNs (20 ng/mL for each, incubated for 24 h), co-cultured with A549, IFNα- and IFNγ (3 h)-pretreated A549 at a 20:1 ratio, and IFNα, IFNγ, and A549 concurrently treated for 24 h were analyzed by flow cytometry to (D and E) detect the activation marker CD107a levels in CD4 + T, CD8 + T cells, and CD45 + CD3 − (nonT) PBMCs (n = 3). CD4 + T and CD8 + T were gated by staining with anti-CD45-Pacific blue, anti-CD3-APC/Cy7, anti-CD8-Alexa488, and anti-CD4-PE. CD107a was detected using anti-CD107a-BV605. (F) In addition, the activation markers IFNG, and cytotoxic marker granzyme B (GZMB) were detected by qPCR in the collected PBMCs after individual treatments. n = 3 and error bars were presented by SD in qPCR analysis. Statistical analysis was achieved by an unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human lung cancer cell lines A549, HepG2, and PLC5 used in this study were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Gene Expression, Expressing, Incubation, RNA sequencing, Cell Culture, Flow Cytometry, Activation Assay, Marker, Staining, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Radiotherapy enhances M1 macrophage immunogenic activity through IFNs induction and stimulation in TP53-wild type tumors

doi: 10.1007/s00262-026-04300-7

Figure Lengend Snippet: Radiotherapy increases IFNs and promotes immunogenic activity but distinct gene expression in the IFNγ- and 8 Gy-treated A549 cells. A A549 cells (5 × 10 5 in a 6-well plate) treated with irradiation (0, 1, 2, 4, 8 Gy) and further incubated for 24 h were collected to detect the p53-downstream gene CDKN1A, MDM2, and tumor marker MKI67 by qPCR. Statistical analysis was achieved by one-way ANOVA. B In addition, IFNA and IFNG and their downstream PD-L1 expression in the irradiated A549 cells and C MDM2 inhibitor Nutlin-3a-treated A549 cells were measured by qPCR. D The healthy PBMCs (1 × 10 6 in a 6-well plate) treated with IFNα and IFNγ (20 ng/mL for each) for 2 h and 24 h were collected and analyzed for the immune activation marker IFNG and cytotoxic marker granzyme B (GZMB) expression using qPCR. E M1 markers TNFA and CXCL10, and M2 markers ARG1 and IL-10 were also investigated in the collected PBMCs with the individual treatments by qPCR. F The healthy PBMCs (1 × 10 6 in a 6-well plate) incubated with irradiation-treated A549 (0, 4, 8 Gy) at a ratio of 20:1 for 24 h were collected and investigated for the immune activation marker IFNG and cytotoxic marker GZMB expression using qPCR. G Meanwhile, M1 markers TNFA, CXCL10, and M2 markers ARG1, IL-10 were also investigated in the collected PBMCs with the individual treatments by qPCR. (H) RNAseq was used to search for the differential genes in the A549 cells (5 × 10 5 in a 6-well plate) treated with IFNγ (20 ng /mL, incubated for 2 h) and x-ray irradiation (8 Gy was selected based on the highest CDKN1A and MDM2 induction, 24 h post-irradiation). There were 75 up-regulated and 12 down-regulated genes selected in IFNγ-treated A549 and 88 up-regulated and 202 down-regulated genes selected in 8 Gy-treated A549 according to log2 fold change > 1 or < -1 with p value < 0.05 (Table S2-4). There was no overlapped gene between IFNγ and 8 Gy treatment. I The 75 and 88 up-regulated genes in IFNγ- and irradiated A549 cells were analyzed by NetworkAnalyst ( https://www.networkanalyst.ca/ ) based on the KEGG dataset, revealing that the differential genes were involved in STATs and p53 signaling pathways, respectively. n = 3 and error bars were presented by SD in qPCR analysis. Statistical analysis was achieved by an unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, non-significant

Article Snippet: The human lung cancer cell lines A549, HepG2, and PLC5 used in this study were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Gene Expression, Irradiation, Incubation, Marker, Expressing, Activation Assay, RNA sequencing, Protein-Protein interactions, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Radiotherapy enhances M1 macrophage immunogenic activity through IFNs induction and stimulation in TP53-wild type tumors

doi: 10.1007/s00262-026-04300-7

Figure Lengend Snippet: Radiotherapy specifically suppresses TP53-wild type tumors. A Flow cytometry based on fluorescent Annexin V staining was used to detect apoptosis in the irradiation (0, 4, and 8 Gy)-treated TP53-wild type A549, HCT116, and TP53null HCT116 cells (5 × 10 5 in a 6-well plate) post 24 h culture. n = 2. B - C qPCR was used to validate the 13 irradiation-mediated genes from RNAseq (Table S3 and Table S4), including the increased MDM2, CYFIP2, STOM, and the decreased MKI67, CENPE, ARGHGAP11A, BRCA1, ASPM, ALAN, TOP2A, FANCI, TOPBP1, and ECT2, in (B) A549 treated with 8 Gy (24 h post-irradiation), C A549 treated with MDM2 inhibitor Nutlin-3a (10 µg/mL for 24 h). D and E qPCR was also used to detect p53-downstream CDKN1A and the selected 13 genes in TP53-wild type and TP53null HCT116 treated with 8 Gy of irradiation (24 h post-irradiation). n = 3 and error bars were presented by SD in qPCR analysis. Statistical analysis was achieved by an unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human lung cancer cell lines A549, HepG2, and PLC5 used in this study were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Flow Cytometry, Staining, Irradiation, RNA sequencing, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Radiotherapy enhances M1 macrophage immunogenic activity through IFNs induction and stimulation in TP53-wild type tumors

doi: 10.1007/s00262-026-04300-7

Figure Lengend Snippet: Radiotherapy induces IFNs in TP53-wild type tumors. A qPCR was used to detect the expression of IFNs in TP53-wild type HepG2, HCT116, TP53-mutant PLC5, and TP53null HCT116 (5 × 10 5 in a 6-well plate) treated with 0, 4, 8 Gy of irradiation (24 h post-irradiation). B The IFNγ-mediated up-regulated genes from RNAseq analysis (Table S2), including PD-L1, ICAM1, BATF, IRF1, SOCS1, HAPLN3, TAP1, PSMB9, and MAFF, were investigated by qPCR in TP53-wild type and TP53null HCT116 (5 × 10 5 in a 6-well plate) treated with 8 Gy of irradiation (24 h post-irradiation). C The cultured medium from the irradiated A549 (0, 4, 8 Gy, post 24 h) was collected to treat parental A549 (5 × 10 5 in a 6-well plate) for 2 h. qPCR was used to measure the selected gene expression. n = 3 and error bars were presented by SD in qPCR analysis. Statistical analysis was achieved by an unpaired two-tailed Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human lung cancer cell lines A549, HepG2, and PLC5 used in this study were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Mutagenesis, Irradiation, RNA sequencing, Cell Culture, Gene Expression, Two Tailed Test

Journal: Natural Products and Bioprospecting

Article Title: Combined treatment with naringin and osthole ameliorates colitis through microbiota–amino acid metabolism and the JNK pathway

doi: 10.1007/s13659-025-00582-z

Figure Lengend Snippet: Combination therapy of naringin and osthole inhibits inflammatory damage in vitro. A – C CCK-8 detection of cell viability. D Perform RT-qPCR on IL-6, IL-1β, TNF-α, and IL-10 in HT-29 cells. E – G Western blotting and quantitative analysis of apoptosis factors Bcl-2, Bax, and Cleaved-Caspase-3. Data are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: The human colon cancer cell line HT-29 was obtained from the American Type Culture Collection (ATCC, USA) and were cultured in DMEM containing 100 IU/mL penicillin, 100 μg/mL streptomycin, and 10% fetal bovine serum (FBS) in a humidified incubator at 37 °C with 5% CO 2 .

Techniques: In Vitro, CCK-8 Assay, Quantitative RT-PCR, Western Blot

Journal: Natural Products and Bioprospecting

Article Title: Combined treatment with naringin and osthole ameliorates colitis through microbiota–amino acid metabolism and the JNK pathway

doi: 10.1007/s13659-025-00582-z

Figure Lengend Snippet: The combined administration of naringin and osthole alleviates colitis in mice by inhibiting the JNK/NF-κB signaling pathway in vitro and in vivo. A , B Western blot for NF-κB signing pathway in vivo and in vitro. C , D Western blot for JNK signing pathway in vivo and in vitro. E Western blot of JNK signing pathway in vitro after JNK inhibitor (SP600125) treatment. Data are presented as mean ± SEM (n = 3). F RT-qPCR of IL-6, IL-1β, TNF-α and IL-10 in HT-29 cells after pretreatment with JNK inhibitor SP600125. Data are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The human colon cancer cell line HT-29 was obtained from the American Type Culture Collection (ATCC, USA) and were cultured in DMEM containing 100 IU/mL penicillin, 100 μg/mL streptomycin, and 10% fetal bovine serum (FBS) in a humidified incubator at 37 °C with 5% CO 2 .

Techniques: In Vitro, In Vivo, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Molidustat Targets a Synthetic Lethal Vulnerability in APC-Mutant Colorectal Cancer through GSTP1 and PHD2 Co-Inhibition

doi: 10.64898/2026.01.31.702998

Figure Lengend Snippet: A. Percentage of Cleaved Caspase 3/7 positive HT29 (top), and RKO (bottom) cells. Cells were treated with Molidustat for 48 hours at indicated concentrations, 10uM Staurosporine was used as a positive control (100% cell death). Mean + SEM is assessed by unpaired two tailed Student’s t-test, **p<0.01, (ns) non-significant. B. Representative images of Cleaved Caspase-3/7 signal in DMSO, Molidustat (90 μM), and Staurosporine treated cells. Scale bar: 300 μm. C. Representative Western Blot of PHD2 levels in HT29 cells. D. Percentage confluency of HT29 cells post-transfection with the indicated guide RNAs. E. Cleaved Caspase-3/7 signal in HT29 cells post-transfection with the indicated crRNAs. Mean + SEM is assessed by two-way ANOVA, *p<0.05. N = 3 biologically independent experiments.

Article Snippet: Human colorectal cancer cell lines HT29 and RKO were obtained from the American Type Culture Collection (ATCC) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Sigma-Aldrich, D6429) supplemented with 10% (v/v) fetal bovine serum (FBS; Gibco, 16000044), 1% (v/v) penicillin-streptomycin (Gibco, 15140122), and 2 mM L-glutamine (Sigma-Aldrich, G7513).

Techniques: Positive Control, Two Tailed Test, Western Blot, Transfection