Journal: Pharmacological reports : PR

Article Title: Anticancer effect of nor-wogonin (5, 7, 8-trihydroxyflavone) on human triple-negative breast cancer cells via downregulation of TAK1, NF-κB, and STAT3

doi: 10.1016/j.pharep.2019.01.001

Figure Lengend Snippet: Proliferation and viability of TNBC cells and non-tumorigenic breast cells after treatment with nor-wogonin and structurally related compounds. A. Chemical structures of nor-wogonin, wogonin, and wogonoside. B. Effects of nor-wogonin (5–80 μM), wogonin (100 μM), and wogonoside (100 μM) on the proliferation of TNBC cells (MDA-MB-231, BT-549, HCC70, and HCC1806) and non-tumorigenic breast cells (MCF-10A and AG11132) were determined by BrdU incorporation assays. Dimethyl sulfoxide (DMSO vehicle) was used as a negative control. C. The cytotoxic effects of nor-wogonin (5–80 μM), wogonin (100 μM), and wogonoside (100 μM) on TNBC cells (MDA-MB-231, BT-549, HCC70, and HCC1806) and non-tumorigenic breast cells (MCF-10A and AG11132) were determined by trypan blue exclusion assays. DMSO was used as a vehicle control. Data are expressed as the means ± SD based on three independent experiments.

Article Snippet: Human TNBC cell lines (MDA-MB-231, BT-549, HCC70, and HCC1806) and non-tumorigenic breast cell line (MCF-10A) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: BrdU Incorporation Assay, Negative Control, Control

Journal: Cancers

Article Title: Significance of LIF/LIFR Signaling in the Progression of Obesity-Driven Triple-Negative Breast Cancer

doi: 10.3390/cancers16213630

Figure Lengend Snippet: Obese-related adiposity conditions enhance LIFR downstream signaling. TNBC cells, BT-549 ( A ), and SUM-159 ( B ) were cultured with ADP-CM for 24 h, and the expression of LIFR target genes was analyzed by RT-qPCR. ( C ) TNBC cells cultured with ADP-CM for 24 h were analyzed using Western blot analysis to measure LIFR downstream signaling proteins. ( D ) The effects of EC359 treatment against adipose conditions on TNBC cell viability were determined using MTT assays. ( E ) The effects of ADP-CM and ADP-CM + EC359 (50 nM) on adiposity-induced cell survival of TNBC cells was measured using colony formation assays. ( F ) Representative images of colonies were shown. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: TNBC cell lines (MDA-MB-231, BT-549, SUM159, and 4T1) were purchased from ATCC and were maintained using ATCC-recommended medium.

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancers

Article Title: Significance of LIF/LIFR Signaling in the Progression of Obesity-Driven Triple-Negative Breast Cancer

doi: 10.3390/cancers16213630

Figure Lengend Snippet: ADP-CM or co-culture of adipocytes enhances LIFR downstream signaling in TNBC and addition of EC359 (100 nM) reduces its activity in vitro. TNBC cells that stably express the STAT3 reporter were used. ADP-CM ( A ) activates STAT3 reporter activity, with EC359 (100 nM) effectively reducing STAT3 activity in ADP-CM conditions. ( B ) BT-549 and MDA-MB-231 cells were cultured with ADP-CM in the presence or absence of EC359 (100 nM) for 24 h, and the expression of LIFR target genes was analyzed by RT-qPCR. ( C ) MDA-MB-231 cells were incubated with or without ADP-CM and EC359 (100 nM) and were analyzed using Western blot analysis to measure LIFR downstream signaling proteins. ( D ) Adipocytes were indirectly co-cultured with MDA-MB-231 cells using a transwell culture system in the presence or absence of EC359 (100 nM) for 24 h. Signaling was profiled by Western blotting. ( E ) Effect of adipose conditions on TNBC cell invasion in the presence or absence of EC359 (100 nM) was determined by Boyden chamber assay and invaded cells were quantified ( F ). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: TNBC cell lines (MDA-MB-231, BT-549, SUM159, and 4T1) were purchased from ATCC and were maintained using ATCC-recommended medium.

Techniques: Co-Culture Assay, Activity Assay, In Vitro, Stable Transfection, Cell Culture, Expressing, Quantitative RT-PCR, Incubation, Western Blot, Boyden Chamber Assay

Journal: Cancers

Article Title: Significance of LIF/LIFR Signaling in the Progression of Obesity-Driven Triple-Negative Breast Cancer

doi: 10.3390/cancers16213630

Figure Lengend Snippet: EC359 reduces tumor volume and weight in adiposity and diet-induced TNBC CDX models. Primary human adipose cells were co-implanted with MDA-MB-231 cells via orthotopic injection (n = 7) and subsequently treated with or without EC359 (5 mg/kg, intraperitoneally, 5 days per week). Tumor volumes ( A ) and tumor weights ( B ) are shown in graph. ( C ) Body weights of Low-fat-diet- and high-fat-diet-induced TNBC xenograft models treated with or without EC359 (5 mg/kg/ip/5 days/week) are shown in graph. Tumor volume ( D ) and tumor weights ( E ) are shown. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: TNBC cell lines (MDA-MB-231, BT-549, SUM159, and 4T1) were purchased from ATCC and were maintained using ATCC-recommended medium.

Techniques: Injection

Journal: Cancers

Article Title: Significance of LIF/LIFR Signaling in the Progression of Obesity-Driven Triple-Negative Breast Cancer

doi: 10.3390/cancers16213630

Figure Lengend Snippet: Low-fat diet and high-fat diet 4T1-TNBC syngeneic model. Diet-induced obesity syngeneic models (n = 8) were treated with vehicle or EC359 (5 mg/kg/ip/5 days/week). ( A ) Body weights shown in graph. Bar graphs of liver ( B ), fat tissue ( C ), spleen ( D ), and tumor volume ( E ) were shown. ( F ) Tumor weights of all the 3 groups are shown. ( G ) Representative images of excised tumors from diet-induced syngeneic xenografts treated with or without EC359. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: TNBC cell lines (MDA-MB-231, BT-549, SUM159, and 4T1) were purchased from ATCC and were maintained using ATCC-recommended medium.

Techniques:

Journal: Cancers

Article Title: Significance of LIF/LIFR Signaling in the Progression of Obesity-Driven Triple-Negative Breast Cancer

doi: 10.3390/cancers16213630

Figure Lengend Snippet: Differential gene expression analysis of TNBC syngeneic xenograft model. ( A ) GSEA reveals pathways with positive enrichment (upregulated pathways) in diet-induced obesity TNBC-4T1 syngeneic xenograft models after HFD feeding. ( B ) GSEA displays pathways with negative enrichment (downregulated pathways) in EC359-treated TNBC-4T1 syngeneic xenograft models after HFD feeding. Gene ontology analysis identifies biological processes with positive enrichment in HFD models ( C ) and negative enrichment in EC359-treated HFD models ( D ). Differentially expressed genes were filtered using log2FoldChange ≥ 2, and p adj < 0.01.

Article Snippet: TNBC cell lines (MDA-MB-231, BT-549, SUM159, and 4T1) were purchased from ATCC and were maintained using ATCC-recommended medium.

Techniques: Gene Expression

Journal: Materials Today Bio

Article Title: A select inhibitor of MORC2 encapsulated by chimeric membranecoated DNA nanocage target alleviation TNBC progression

doi: 10.1016/j.mtbio.2025.101497

Figure Lengend Snippet: MORC2 is highly expressed in TNBC and is closely associated with poor biological behavior (high proliferation and invasion ability) and prognosis. (A) Kaplan-Meier survival curve for patients with high/low expression of MORC2. Data come from the KM-plotter database. (B) Immunofluorescence showed that the expression of MORC2 is significantly higher in TNBC tissue than adjacent nontumor tissue (scale = 100 μm). (C) Western blot results of MORC2 expression in TNBC tissue or adjacent nontumor tissue. (D) Western blot results of MORC2 expression in normal breast epithelial cell line (MCF-10A) or TNBC cell lines (MDA-MB-231, BT-549 and MDA-MB-468). (E) Knockdown of MORC2 decreased the proliferation capability of both MDA-MB-231 and BT-549. (F) Knockdown of MORC2 decreased the invasion capability of both MDA-MB-231 and BT-549. (G) Cell cycle changed after Knockdown of MORC2 in MDA-MB-231 and BT-549 cells. (H) Growth curves of inoculated xenograft tumors in shMORC2 or shNC groups. ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, no significance.

Article Snippet: Moreover, human TNBC cell lines (MDA-MB-231, MDA-MB-468 and BT-549) and normal breast epithelial cell line (MCF-10A) were purchased from Procell (Wuhan, China).

Techniques: Expressing, Immunofluorescence, Western Blot, Knockdown

Journal: Materials Today Bio

Article Title: A select inhibitor of MORC2 encapsulated by chimeric membranecoated DNA nanocage target alleviation TNBC progression

doi: 10.1016/j.mtbio.2025.101497

Figure Lengend Snippet: Angoline has strong binding capacity with MORC2 protein and causes TNBC cell death by inducing apoptosis. (A) Diagram of the chemical structure of Angoline. (B) Molecular docking software predicts binding sites, interaction forces and distances between Angoline and MORC2 protein. ASP, asparticacid; ASN, asparagine; MET, methionine; LYS, lysine; SER, serine; VAL, valine; LEU, leucine. (C) In vitro molecular interaction experiment showed that the binding strength between MORC2 protein and Angoline varies with the concentration of Angoline. (D) The binding force between MORC2 protein and Angoline increased with the mutation of amino acids at the binding site. (E) The killing effect of Angoline on TNBC cells was enhanced with increasing concentration. (F) The killing effect of Angoline on TNBC cells was reduced by apoptosis inhibitor (Z-VAD-FMK). (G) The apoptosis-inducing effect of Angoline on TNBC cells was weakened by apoptosis inhibitor (Z-VAD-FMK). (H) YO-PRO-1 staining showed that the pro-apoptotic effect of angoline on TNBC cells can be attenuated by apoptosis inhibitor (Z-VAD-FMK) ((bar, 100 μm)). ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, no significance.

Article Snippet: Moreover, human TNBC cell lines (MDA-MB-231, MDA-MB-468 and BT-549) and normal breast epithelial cell line (MCF-10A) were purchased from Procell (Wuhan, China).

Techniques: Binding Assay, Software, In Vitro, Concentration Assay, Mutagenesis, Staining

Journal: Communications Biology

Article Title: LIFR inhibition enhances the therapeutic efficacy of HDAC inhibitors in triple negative breast cancer

doi: 10.1038/s42003-021-02741-7

Figure Lengend Snippet: a TNBC model cells (MDA-MB-231 and BT-549) were treated with indicated HDACi (vorinostat: 10 µM; panobinostat: 1 µM; romidepsin: 1 µM; givinostat: 1 µM) for 24 h and expression of LIFR, p-STAT3(Y705), and STAT3 were determined using Western blotting. b MDA-MB-231 and BT-549 cells were treated with vorinostat (10 µM) for 10 h and levels of LIFR were measured by RT-qPCR. RT-qPCR data were normalized to GAPDH and data are representative of three independent experiments ( n = 3). c MDA-MB-231 and BT-549 cells stably expressing STAT3-luc reporter were treated with indicated HDACi and reporter activity was measured after 24 h. Data are representative of three independent experiments ( n = 3). d BT-549 vec or BT-549 LIFR-KO cells were treated with indicated HDACi (vorinostat: 10 µM; panobinostat: 1 µM, romidepsin: 1 µM) for 10 h and induction of LIFR, LIF, and p-STAT3(Y705) was measured by Western blotting. The effect of LIFR-KO on the activity of HDACi was determined using MTT cell viability assay ( e ) and clonogenic survival assay ( f ). e , f Data are representative of three independent experiments ( n = 3). Error bars represent SD. In b , e , and f , p -values were calculated using two-way ANOVA. In c , p -values were calculated using one-way ANOVA.

Article Snippet: Human TNBC cells (MDA-MB-231, BT-549, MDA-MB-468, HCC1806, and HCC70) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured as per ATCC guidelines.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Stable Transfection, Activity Assay, Viability Assay, Clonogenic Cell Survival Assay

Journal: Communications Biology

Article Title: LIFR inhibition enhances the therapeutic efficacy of HDAC inhibitors in triple negative breast cancer

doi: 10.1038/s42003-021-02741-7

Figure Lengend Snippet: MDA-MB-231 and BT-549 cells were treated with indicated concentrations of vorinostat ( a ), for 72 h in the presence or absence of EC359 (MDA-MB-231: 5 nM; BT-549: 10 nM) and the cell viability was measured by MTT assay ( n = 3). Combination Index (CI) values with respect to different concentrations were shown in the bottom of each graph. b Effect of EC359 + HDACi combination therapy on the cell survival of TNBC cells was measured using colony formation assays ( n = 3). Representative images from three independent experiments are shown on the left panel, and quantitation of colonies is presented on the right panel. c Effect of EC359 + HDACi combination therapy on cell invasion of MDA-MB-231 and BT-549 cells was determined using matrigel invasion chamber assays. Data are representative of three independent experiments ( n = 3). Representative images of invaded cells are shown and the number of invaded cells were quantitated ( d ). e Effect of EC359 + HDACi therapy on apoptosis was determined using Annexin V-PI staining and Caspase 3/7 activity (Caspase-Glo 3/7® assay) in MDA-MB-231 cells. Data are representative of three independent experiments ( n = 3). Error bars represent SD. The combination index (CI) of EC359 + HDACi therapy was determined using Chou-Talalay method. p -values were calculated using two-way ANOVA. **** P < 0.0001.

Article Snippet: Human TNBC cells (MDA-MB-231, BT-549, MDA-MB-468, HCC1806, and HCC70) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured as per ATCC guidelines.

Techniques: MTT Assay, Quantitation Assay, Staining, Activity Assay, Caspase-Glo Assay

Journal: Communications Biology

Article Title: LIFR inhibition enhances the therapeutic efficacy of HDAC inhibitors in triple negative breast cancer

doi: 10.1038/s42003-021-02741-7

Figure Lengend Snippet: a Breast tissue microarray consisting of normal adjacent tissue (NAT, n = 6), benign ( n = 6), ER + PR + ( n = 50), ER + HER2+ ( n = 11), HER2+ ( n = 30), and TNBC ( n = 61) samples were subjected to immunohistochemical staining using LIFR antibody and the intensity and positivity of LIFR staining was quantitated. Scale bar represents 100 µm. b MDA-MB-231 ( n = 4) and c MDA-MB-468 ( n = 6) xenografts in SCID mice were treated with vehicle or EC359 (5 mg/kg/day) or vorinostat (100 mg/kg/day) or in combination. Tumor volume and weight of vehicle and treated tumors were measured. d the proliferation of MDA-MB-468 xenograft tumors was determined using Ki67 immunostaining. Representative Ki67 staining from each treatment condition is shown in the upper panel and quantification plot is shown in the lower panel. In a , p -value was calculated using one-way ANOVA. Error bars represent SD. In b , c , and d , p -values were calculated using t test and two-way ANOVA.

Article Snippet: Human TNBC cells (MDA-MB-231, BT-549, MDA-MB-468, HCC1806, and HCC70) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured as per ATCC guidelines.

Techniques: Microarray, Immunohistochemical staining, Staining, Immunostaining