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: Breast Cancer Research : BCR

Article Title: Exosome-delivered METTL14 drives hypoxia-induced proliferation, metastasis, and glycolysis of breast cancer cells through regulating TRIM16-mediated FGF7 ubiquitination

doi: 10.1186/s13058-025-02099-2

Figure Lengend Snippet: Expression patterns of FGF7 in TNBC tissues and cells. ( A ) RT-qPCR assay was used to detect the expression level of FGF7 in 46 pairs of TNBC tumor tissues and adjacent normal tissues (Unpaired t-test). ( B ) The overall survival of TNBC patients with FGF7 high or low levels was determined by Kaplan-Meier analysis (Unpaired t-test). ( C ) FGF7 expression was measured in BC patients in I + II stage ( N = 19) and III stage ( N = 27) using RT-qPCR assay (Unpaired t-test). ( D ) RT-qPCR assay was employed to assess the expression of FGF7 in BC patients with ( N = 28) or without ( N = 18) lymph node metastasis (Unpaired t-test). ( E ) Western blot assay was applied to determine the protein level of FGF7 in BC tumor tissues and adjacent normal tissues (Unpaired t-test). ( F ) FGF7 protein level in MCF-10 A cells and TNBC cell lines (MDA-MB-468, BT-549, and MDA-MB-231) was determined using Western blot assay (one-way ANOVA). ( G ) FGF7 protein level was examined in TNBC cells (BT-549 and MDA-MB-231) under hypoxia at time points (0 h, 12 h, 24 h, 48 h) using western blot assay (two-way ANOVA). N = 3

Article Snippet: TNBC cell lines (MDA-MB-468, CL-0290; BT-549, CL-0041; MDA-MB-231, CL-0150) and normal human breast epithelial cell line (MCF-10 A, CL-0525) were respectively incubated in medium (CM-0290B, CM-0041, CM-0150B, CM-0525, Procell, Wuhan, China) at 37 ̊C with 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Exosome-delivered METTL14 drives hypoxia-induced proliferation, metastasis, and glycolysis of breast cancer cells through regulating TRIM16-mediated FGF7 ubiquitination

doi: 10.1186/s13058-025-02099-2

Figure Lengend Snippet: Effects of FGF7 knockdown on TNBC cell proliferation, metastasis, and glycolysis under hypoxia. ( A-G ) BT-549 and MDA-MB-231 cells were treated with normoxia (Normal), Hypoxia, Hypoxia + si-NC, or Hypoxia + si-FGF7. ( A ) FGF7 protein level was evaluated in treated TNBC cells using western blot. ( B ) Cell viability was determined using MTT assay. ( C and D ) EdU-positive cells were detected using EdU assay. ( E ) Cell invasion was assessed using Transwell assay. ( F and G ) Cell migration ability was measured using wound healing assay. ( H ) Cell viability was examined in BT-549 and MDA-MB-231 cells treated with normoxia (Normal), Hypoxia, Hypoxia + 2-DG (Glycolysis inhibitor). ( I-K ) The glucose consumption, lactic acid production, and ATP/ADP ratios were assessed in BT-549 and MDA-MB-231 cells treated with normoxia (Normal), Hypoxia, Hypoxia + si-NC, or Hypoxia + si-FGF7 using relevant kits. Two-way ANOVA, N = 3

Article Snippet: TNBC cell lines (MDA-MB-468, CL-0290; BT-549, CL-0041; MDA-MB-231, CL-0150) and normal human breast epithelial cell line (MCF-10 A, CL-0525) were respectively incubated in medium (CM-0290B, CM-0041, CM-0150B, CM-0525, Procell, Wuhan, China) at 37 ̊C with 5% CO 2 .

Techniques: Knockdown, Western Blot, MTT Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay

Journal: Breast Cancer Research : BCR

Article Title: Exosome-delivered METTL14 drives hypoxia-induced proliferation, metastasis, and glycolysis of breast cancer cells through regulating TRIM16-mediated FGF7 ubiquitination

doi: 10.1186/s13058-025-02099-2

Figure Lengend Snippet: TRIM16/FGF7 affected TNBC cell proliferation, metastasis, and glycolysis under hypoxia. ( A ) FGF7 protein level was determined in BT-549 and MDA-MB-231 cells transfected with OE-NC or OE-FGF7 using western blot assay. ( B-J ) BT-549 and MDA-MB-231 cells were treated with normoxia (Normal), Hypoxia, Hypoxia + OE-NC, Hypoxia + OE-TRIM16, or Hypoxia + OE-TRIM16 + OE-FGF7. ( B-D ) Cell proliferative ability was assessed using MTT and EdU assays. ( E-G ) Cell invasion and migration were measured using Transwell and wound healing assays. ( H-J ) The glucose consumption, lactic acid production, and ATP/ADP ratios were detected using relevant kits. Two-way ANOVA, N = 3

Article Snippet: TNBC cell lines (MDA-MB-468, CL-0290; BT-549, CL-0041; MDA-MB-231, CL-0150) and normal human breast epithelial cell line (MCF-10 A, CL-0525) were respectively incubated in medium (CM-0290B, CM-0041, CM-0150B, CM-0525, Procell, Wuhan, China) at 37 ̊C with 5% CO 2 .

Techniques: Transfection, Western Blot, Migration

Journal: Breast Cancer Research : BCR

Article Title: Exosome-delivered METTL14 drives hypoxia-induced proliferation, metastasis, and glycolysis of breast cancer cells through regulating TRIM16-mediated FGF7 ubiquitination

doi: 10.1186/s13058-025-02099-2

Figure Lengend Snippet: METTL14/TRIM16 regulated hypoxia-mediated TNBC cell proliferation, metastasis, and glycolysis. ( A ) TRIM16 protein level was measured in BT-549 and MDA-MB-231 cells transfected with si-NC or si-TRIM16 using western blot assay. ( B-K ) BT-549 and MDA-MB-231 cells were treated with normoxia (Normal), Hypoxia, Hypoxia + si-NC, Hypoxia + si-METTL14, or Hypoxia + si-METTL14 + si-TRIM16. ( B-D ) MTT and EdU assays were performed to measure cell proliferative ability. ( E-G ) Transwell and wound healing assays were employed to detect cell invasion and migration. ( H-J ) Relevant kits analyzed the glucose consumption, lactic acid production, and ATP/ADP ratios. ( K ) FGF7 protein level was examined using western blot assay. Two-way ANOVA, N = 3

Article Snippet: TNBC cell lines (MDA-MB-468, CL-0290; BT-549, CL-0041; MDA-MB-231, CL-0150) and normal human breast epithelial cell line (MCF-10 A, CL-0525) were respectively incubated in medium (CM-0290B, CM-0041, CM-0150B, CM-0525, Procell, Wuhan, China) at 37 ̊C with 5% CO 2 .

Techniques: Transfection, Western Blot, Migration

Journal: Breast Cancer Research : BCR

Article Title: Exosome-delivered METTL14 drives hypoxia-induced proliferation, metastasis, and glycolysis of breast cancer cells through regulating TRIM16-mediated FGF7 ubiquitination

doi: 10.1186/s13058-025-02099-2

Figure Lengend Snippet: Hypoxia-induced exosomal transport of METTL14 in TNBC cells. ( A ) The representative micrograph of round-shaped vesicles by TEM (scale bars = 200 nm). ( B ) Western blot analysis of exosome-specific markers (CD63, CD81, and TSG101) in their corresponding exosomes from BT-549 and MDA-MB-231 cells. ( C and D ) METTL14 mRNA and protein expression were determined in exosomes from BT-549 and MDA-MB-231 treated with normoxia (Normal) or Hypoxia using RT-qPCR and western blot assays. ( E ) Representative fluorescent images of BT-549 and MDA-MB-231 cells co-incubated with Hypoxia-induced exosomes from the corresponding TNBC cells. ( F ) METTL14 protein level was determined in BT-549 and MDA-MB-231 cells co-incubated with PBS or Hypoxic-treated exo using western blot assay. Two-way ANOVA, N = 3

Article Snippet: TNBC cell lines (MDA-MB-468, CL-0290; BT-549, CL-0041; MDA-MB-231, CL-0150) and normal human breast epithelial cell line (MCF-10 A, CL-0525) were respectively incubated in medium (CM-0290B, CM-0041, CM-0150B, CM-0525, Procell, Wuhan, China) at 37 ̊C with 5% CO 2 .

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Incubation

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