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: Journal of Cellular and Molecular Medicine

Article Title: Aberrant methylation of WD‐repeat protein 41 contributes to tumour progression in triple‐negative breast cancer

doi: 10.1111/jcmm.15344

Figure Lengend Snippet: Expression pattern of WDR41 in human breast cancer cells and tissues (A) Expression of WDR41 in breast cancer cell lines was evaluated by Western blotting. MDA‐MB‐231, BT549, MDA‐MB‐468 and MDA‐MB‐453 were the TNBC cell lines; MCF‐10A and MCF‐7 were the normal breast cell line and low invasive capability cells, respectively. The SKBR3 cell line was ER‐ and PR‐negative, but Her2‐positive. B, The mRNA level of WDR41 was determined by qRT‐PCR in normal mammary epithelial cells (MCF‐10A) and breast tumour cells (MCF‐7, MDA‐MB‐231 and SKBR3). C, Immunohistochemical staining of WDR41 in human breast cancer samples and corresponding adjacent tissues. Scale bar: 20 μm. D, Protein expression of WDR41 in human breast cancer tissues and paired para‐carcinoma tissues was measured by Western blotting. E, Relative protein expression of WBP2 in panel D. F, The mRNA level of WDR41 in human breast cancer tissues and its paired para‐carcinoma tissues was measured by qRT‐PCR. G, Immunohistochemical staining with WDR41‐specific antibody in different pathological grades of human breast cancer tissue chips. Scale bar: 50 μm. P, para‐carcinoma, T, tumour. * P < .05, ** P < .01. ER, oestrogen receptor; PR, progesterone receptor; qRT‐PCR, quantitative real‐time PCR; TNBC, triple‐negative breast cancer; WDR41, WD‐repeat protein 41

Article Snippet: The normal mammary epithelial cell line (MCF‐10A), oestrogen receptor (ER)‐positive breast cancer cell (MCF‐7), TNBC cell lines (MDA‐MB‐231, BT549, MDA‐MB‐468 and MDA‐MB‐453) and ER/progesterone receptor (PR)‐positive and Her2‐negative breast cancer cell lines (SKBR3 and HS578t, respectively) were purchased from American Type Culture Collection.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemical staining, Staining, Real-time Polymerase Chain Reaction

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: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circRNA expression profiles in TNBC. a. The cluster heat map showed the differentially expressed circRNAs over 3-fold change. Red color indicates high expression level, and green color indicates low expression level. The black arrow indicates circGFRA1. b. The scatter plot was used for assessing the variation in circRNA expression between TNBC cell lines and HME cell line. The values of x and y axes in the scatter plot were the normalized signal values of the samples (log2 scaled). The green lines are fold-change lines. c. The top 10 upregulated circRNAs were shown. d. qRT-PCR was performed to verify the expression of the top eight upregulated circRNAs. All the data are shown as the mean ± s.e.m.

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 is upregulated and correlated with poor clinical outcomes in TNBC. a. The expression level of circGFRA1 was determined by qRT-PCR in 11 different mammary cell lines, including two HME cell line (MCF-10A and 184A1) and nine breast cancer cell lines. circGFRA1 expression was normalized using β-actin expression. b. The expression level of circGFRA1 in 51 TNBC tissues and their matched normal adjacent tissues was determined by qRT-PCR. c. OS curves for 222 TNBC patients with high or low circGFRA1 expression. d. DFS curves for 222 TNBC patients with high or low circGFRA1 expression. All the data are shown as the mean ± s.e.m.

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 serves as a sponge for miR-34a. a. The levels of nuclear control transcript (U6), cytoplasmic control transcript (GAPDH mRNA) and circGFRA1 were assessed by qRT-PCR in nuclear and cytoplasmic fractions. b. The predicted binding sites of miR-34a within circGFRA1 were shown. c. Cells were transfected with miR-34a mimic or miR-34a inhibitor, qRT-PCR analysis demonstrated that the transfection was successful. d. Luciferase assay of MDA-MB-231 cells cotransfected with miR-34a mimics or miR-34a inhibitor and luciferase reporter containing circGFRA1 3′-UTR (circGFRA1 wt) or mutant construct (circGFRA1 mut). e. Cells were transfected as described, and the expression of circGFRA1 was determined by qRT-PCR. f. Cells were transfected with si-NC or si-circGFRA1, and the expression of miR-34a was determined by qRT-PCR. U6 snRNA was used as an internal control. g. MS2-based RIP assay in TNBC cells transfected with MS2bs-circGFRA1, MS2bs-circGFRA1mt, or MS2bs-Rluc (control vector) ( n = 3). All the data are shown as the mean ± s.e.m., * P < 0.05 and ** P < 0.01

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Control, Quantitative RT-PCR, Binding Assay, Transfection, Luciferase, Mutagenesis, Construct, Expressing, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a

doi: 10.1186/s13046-017-0614-1

Figure Lengend Snippet: circGFRA1 and GFRA1 act as ceRNAs in TNBC through regulation of miR-34a. a. The expression level of GFRA1 was determined by qRT-PCR (left) and western blot (right) in 11 mammary cell lines. β-actin was used as a control. b. The expression level of GFRA1 in 51 TNBC tissues and their matched normal adjacent tissues was determined by qRT-PCR. c. The predicted binding sites of miR-34a within GFRA1 were shown. d. Luciferase assay of MDA-MB-231 cells cotransfected with miR-34a mimics or miR-34a inhibitor and luciferase reporter containing GFRA1 3′-UTR (GFRA1 wt) or mutant construct (GFRA1 mut). e. Cells were transfected as described, and the expression of GFRA1 was determined by qRT-PCR. f. Cells were transfected as described, and the expression of GFRA1 was determined by western blot analysis. G. Cells were transfected with si-NC or si-GFRA1, and the expression of miR-34a was determined by qRT-PCR. h. Cells were transfected with si-NC, si-circGFRA1 or si-circGFRA1 + miR-34a inhibitor, and the expression of GFRA1 was determined by qRT-PCR. i. Cells were transfected with si-NC, si-GFRA1 or si-GFRA1 + miR-34a inhibitor, and the expression of circGFRA1 was determined by qRT-PCR. j. Cells were transfected with miR-NC, miR-34a or si-circGFRA1 + miR-34a, and CCK8 assay was performed to assess cell proliferation. k. Cells were transfected as described, and apoptosis assay was performed after transfection. All the data are shown as the mean ± s.e.m., * P < 0.05 and ** P < 0.01

Article Snippet: Three TNBC cell lines (MDA-MB-231, BT549 and MDA-MB-468) and normal mammary epithelial cell line (MCF-10A) were analyzed by Arraystar Human circRNA Array V2.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Binding Assay, Luciferase, Mutagenesis, Construct, Transfection, CCK-8 Assay, Apoptosis Assay