Journal: Hormones & Cancer

Article Title: Aryl Hydrocarbon Receptor in Breast Cancer—A Newly Defined Prognostic Marker

doi: 10.1007/s12672-013-0160-z

Figure Lengend Snippet: MCF-7, T47D, ZR75-1, and MDA-MB-231 were all treated with 10 nM TCDD for 24 h in phenol red- and FBS-free medium. a, b Ki-67 LI of both ER-positive and ER-negative breast cancer cell lines decreased by TCDD administration. Bars mean ± SE (n = 3). *P < 0.05, **P < 0.01 compared with control. c CYP1A1 was induced following TCDD administration, consistent with AhR activation by TCDD. Bars mean ± SE (n = 3). ***P < 0.001 compared with control. Scale bars 100 μm

Article Snippet: Breast Carcinoma Cell Line and Cell Culture Human breast carcinoma cell lines MCF-7, T47D, ZR75-1, and MDA-MB-231 were all obtained from Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Activation Assay

Journal: Hormones & Cancer

Article Title: Aryl Hydrocarbon Receptor in Breast Cancer—A Newly Defined Prognostic Marker

doi: 10.1007/s12672-013-0160-z

Figure Lengend Snippet: a Wound healing assay in T47D and ZR75-1. T47D were exposed to 10 nM TCDD, 10 nM estrogen (E2) and/or 10 μM AhR antagonist (CH-223191), and ZR75-1 were exposed to 10 nM E2 and/or 10 nM TCDD in phenol red- and FBS-free medium for 24 h. Exposure to 10 nM E2 in serum-free medium for 24 h significantly induced migration of T47D and ZR75-1. A 10 nM TCDD administration significantly inhibited E2-dependent induction of cell migration in both cell lines. The inhibitory effects of TCDD on E2-dependent cell migration were significantly reduced by specific AhR antagonist in T47D cells. Bars mean ± SE (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001. b We examined the effects of AhR on cell invasion using Matrigel-coated transwell chambers. T47D and ZR75-1 were treated with 10 nM E2 and/or 10 nM TCDD in phenol red- and FBS-free medium. After 24 h, the cells were plated in the upper chambers (5 × 105 cells/ml). Phenol red-free charcoal-stripped RPMI 1640 medium with 10 % FBS was used as a chemoattractant in the lower chamber. After 24 h, the cells invaded were counted. With 10 nM E2 administration, T47D and ZR75-1 invaded with forming larger clusters, and the number of invaded cells counted at three to five random fields in hotspot areas was significantly higher than that in control. A 10 nM TCDD administration significantly suppressed E2-dependent induction of cell invasion of T47D (P = 0.0431) and ZR75-1 (P = 0.0452). Bars mean ± SE (n = 3–5). *P < 0.05, **P < 0.01. c Cell proliferation assay using WST-8 method. Both T47D and ZR75-1 demonstrated no significant changes of the number of the cells between control and 10 nM E2 administration in serum-free medium for 24 h. Bars mean ± SE (n = 6). d We confirmed the activation of AhR by examining CYP1A1 induction. Bars mean ± SE (n = 3). ***P < 0.001 compared with control

Article Snippet: Breast Carcinoma Cell Line and Cell Culture Human breast carcinoma cell lines MCF-7, T47D, ZR75-1, and MDA-MB-231 were all obtained from Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Wound Healing Assay, Migration, Proliferation Assay, Activation Assay