Journal: Frontiers in Oncology

Article Title: Carnosol, a Natural Polyphenol, Inhibits Migration, Metastasis, and Tumor Growth of Breast Cancer via a ROS-Dependent Proteasome Degradation of STAT3

doi: 10.3389/fonc.2019.00743

Figure Lengend Snippet: Carnosol inhibits the STAT3 signaling pathway. (A) Concentration-dependent decrease of phospho-STAT3 and STAT3 protein in carnosol-treated MDA-MB-231 cells. Cells were treated with vehicle (DMSO) or indicated concentrations of carnosol for 24 h, then whole-cell extracts were subjected to Western blot analysis for the phosphorylated and non-phosphorylated form of STAT3 and for β-actin (loading control). (B) Carnosol decreases the level of STAT3 protein in Hs578T, MCF-7, and T47D breast cancer cell lines. Whole cell lysates from different breast cancer cell lines treated with the indicated concentrations of carnosol were resolved on 8% SDS-PAGE and analyzed by western blot for STAT3 protein. (C) Western blot analysis of STAT3 protein level in HCT116 colorectal cancer cells. Western blotting was performed as described above.

Article Snippet: Human breast cancer cells MDA-MB-231(cat# 300275), MCF-7 (cat# 300273) and T47D (cat# 300353) were purchased from Cell Line Service (CLS)-GmbH.

Techniques: Concentration Assay, Western Blot, Control, SDS Page

Journal: PLOS One

Article Title: Ivermectin inhibits ER, HER2, and TGF-β pathways in ER-positive and endocrine-resistant breast cancer cells

doi: 10.1371/journal.pone.0348260

Figure Lengend Snippet: MTT assay measured cell viability of ER-positive breast cancer cells: (A) MCF-7 and (B) T-47D, were treated with (+E 2 ) or without (−E 2 ) 17β-estradiol at physiologic level (10 nM E 2 ) in the presence of nontoxic concentrations of IVM (1, 3, 5 μM) or 4-OHT (5, 10 μM) as a positive control for 5 days. (C–E) The inhibitory effect of IVM on ERα and HER2 protein levels of ER-positive breast cancer was performed after the treatments with various concentrations of IVM with (+E 2 ) or without (−E 2 ) 17β-estradiol (10 nM E 2 ) for 24 h in MCF-7. (F) mRNA expression of the ESR1 gene (ERα) was determined by qPCR analysis in MCF-7 cells. The graphs showed the mean ± SEM. The blue and red asterisks highlight the significant differences in comparison to non-treatment without (−E 2 ) and with (+E 2 ), respectively. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the non-treatment control (n = 3).

Article Snippet: The ER-positive breast cancer cell lines: MCF-7 and T-47D, along with the tamoxifen-resistant T-47D Tam1, were obtained from the American Type Culture Collection: ATCC (Virginia, USA).

Techniques: MTT Assay, Positive Control, Expressing, Comparison, Control

Journal: PLOS One

Article Title: Ivermectin inhibits ER, HER2, and TGF-β pathways in ER-positive and endocrine-resistant breast cancer cells

doi: 10.1371/journal.pone.0348260

Figure Lengend Snippet: MTT assay measured cell viability of ER-positive breast cancer cells: (A) MCF-7 and (B) T-47D, were treated with (+E 2 ) or without (−E 2 ) 17β-estradiol at physiologic level (10 nM E 2 ) in the presence of nontoxic concentrations of IVM (1, 3, 5 μM) or 4-OHT (5, 10 μM) as a positive control for 5 days. (C–E) The inhibitory effect of IVM on ERα and HER2 protein levels of ER-positive breast cancer was performed after the treatments with various concentrations of IVM with (+E 2 ) or without (−E 2 ) 17β-estradiol (10 nM E 2 ) for 24 h in MCF-7. (F) mRNA expression of the ESR1 gene (ERα) was determined by qPCR analysis in MCF-7 cells. The graphs showed the mean ± SEM. The blue and red asterisks highlight the significant differences in comparison to non-treatment without (−E 2 ) and with (+E 2 ), respectively. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus the non-treatment control (n = 3).

Article Snippet: The ER-positive breast cancer cell lines: MCF-7 and T-47D, along with the tamoxifen-resistant T-47D Tam1, were obtained from the American Type Culture Collection: ATCC (Virginia, USA).

Techniques: MTT Assay, Positive Control, Expressing, Comparison, Control

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 depletion in breast tumor cells increases lymph node adhesion and cell migration toward VN. a NDRG4 expression meta-analysis in human breast cancer cell lines using GOBO application . Breast cancer subtypes can be identified by different colors: red = Basal A, gray = Basal B and blue = Luminal cells. b Analysis of NDRG4 mRNA expression levels in MCF-7 cells transfected with NDRG4-shRNAs or scramble control (shSCR). Expression levels are relative to wild type cells and normalized to hydroxymethylbilane synthase (HMBS) gene. Error bars represent SEM of biological replicates ( n = 5). *** p < 0.001, ns = not significant, by one-way ANOVA. c Western blot analysis of NDRG4 expression in cytoplasmic extracts of MCF-7 shNDRG4s or shSCRs cells, using antibodies against NDRG4 and Actin. MCF-7 cells express all the three isoforms of NDRG4: 37 kDa (NDRG4-B), 39 kDa (NDRG4-B var ) and 41 kDa isoform (NDRG4-H). For simplicity, results obtained for two independent shNDRG4 clones were grouped and presented as the shNDRG4 group. ( d , left) Representative images of adherent red fluorescent MCF-7 cells on frozen rat lymph node sections. d , e Quantification of adherent MCF7 ( d ) and T47D ( e ) breast tumor cells in lymph nodes sections (right, n = 4 independent experiments for MCF-7 and 3 for T47D, ** p < 0.01 by two-tailed t test). f Stationary adhesion assays showed that NDRG4 knockdown promotes ‘adhesive switch’ between FN and VN. Error bars represent SEM of biological replicates ( n = 4). Cells do not adhere to albumin control (BSA, data not shown). * p < 0.01, ** p < 0.01, ns = not significant by two-tailed t tests. g , h Haptotactic cell migration toward VN was analyzed by transwell migration assay in MCF-7 ( g ) and T47D ( h ) cell lines. Error bars represent SEM of biological replicates ( n = 4). *** p < 0.001 by two-tailed t tests

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: Migration, Expressing, Transfection, Western Blot, Clone Assay, Two Tailed Test, Transwell Migration Assay

Journal: NPJ Breast Cancer

Article Title: NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

doi: 10.1038/s41523-019-0106-x

Figure Lengend Snippet: NDRG4 knockdown promotes clustering of β1-integrin at the leading edge of T47D cells. Representative confocal images of β1 integrin subunit (MAB1965, green) at the ventral cell surface of VN-adherent T47D shNDRG4 or shSCR cells

Article Snippet: For stable transfection of NDRG4 shRNA into MCF-7 and T47D breast tumor cells, the pGFP-V-RS vector containing shRNA inserts which targeted the 3′-unttranslated region of NDRG4 (TRCN0000134583 and TRCN0000137216) was purchased (Origene).

Techniques: