Journal: Journal of the Endocrine Society

Article Title: Riluzole Suppresses Growth and Enhances Response to Endocrine Therapy in ER+ Breast Cancer

doi: 10.1210/jendso/bvad117

Figure Lengend Snippet: Single-agent riluzole inhibits tumor growth in vivo, but the combination with fulvestrant is not better than fulvestrant alone in the HCI-013EI ILC PDX model. A, Forty-eight mice were orthotopically implanted with a 1- to 3-mm 3 HCI-013EI PDX fragment without E2 supplementation, then followed until tumors reached ∼100 mm 3 before enrollment into 1 of 4 treatment arms: control (n = 5), fulvestrant (n = 5), riluzole (n = 5), or the combination (n = 5). Mice were monitored for tumor growth (measured by calipers) and body weight twice per week. Data are presented as mean tumor volume ± SEM, and were analyzed by mixed-effects analysis followed by Dunnett's multiple comparisons tests at each timepoint vs control. B, At the end of the study, tumors were collected and weighed. The graph illustrates the summary of the collected data, which were analyzed using Browne-Forsyth and Welch ANOVA followed by Dunnett's T3 multiple comparisons tests. C, Graph showing relative tumor size at endpoint according to RECIST 1.1 criteria. It shows that 2 of 5 tumors in the fulvestrant group and 3 of 5 in the combination group achieved partial response. Abbreviations in Figure 7C graph: PD, progressive disease; PR, partial response; SD, stable disease. D and E, The tumors collected from each treatment group were formalin-fixed, paraffin-embedded, sectioned, and stained with proliferating cell nuclear antigen (PCNA) and Caspase-3 by IHC. These antibodies served as a proxy for proliferation and apoptosis, respectively. The stained samples were analyzed, and the data were presented graphically in D for PCNA, and E for Caspase-3.

Article Snippet: PDEs were treated with 100nM fulvestrant, 10μM riluzole, the combination, or solvent control (DMSO) for 48 hours before formalin fixation, paraffin embedding, sectioning, and staining for PCNA (1:1000, Santa Cruz Biotechnology Cat# sc-56, RRID:AB_628110), cleaved caspase 3 (1:300, Cell Signaling Technology Cat# 9661, RRID:AB_2341188), and Ki67 (1:500, Abcam Cat# ab16667, RRID:AB_302459).

Techniques: In Vivo, Control, Formalin-fixed Paraffin-Embedded, Staining

Journal: Journal of the Endocrine Society

Article Title: Riluzole Suppresses Growth and Enhances Response to Endocrine Therapy in ER+ Breast Cancer

doi: 10.1210/jendso/bvad117

Figure Lengend Snippet: Riluzole plus fulvestrant significantly inhibits proliferation in primary breast tumor explant cultures. A, Pathologic data for 5 patient-derived explants (PDEs). ER, PR, and Ki67% are from the initial surgical specimen, and NOS = not otherwise specified. *denotes the PDE for which representative images are shown in panel C. B, PDEs were treated with 100nM fulvestrant, 10μM riluzole, the combination, or DMSO control (vehicle) for 2 days prior to formalin fixation, paraffin embedding, sectioning, and staining for PCNA by IHC. Data are presented as change relative to vehicle (set to 0) for each explant and analyzed by one-sample t test vs 0 (vehicle). * P = .013 Vehicle vs Combination. *denotes the PDE for which representative images are shown in panel C. C, Representative images of PCNA and Caspase-3 staining from PDE #1055 (ILC).

Article Snippet: PDEs were treated with 100nM fulvestrant, 10μM riluzole, the combination, or solvent control (DMSO) for 48 hours before formalin fixation, paraffin embedding, sectioning, and staining for PCNA (1:1000, Santa Cruz Biotechnology Cat# sc-56, RRID:AB_628110), cleaved caspase 3 (1:300, Cell Signaling Technology Cat# 9661, RRID:AB_2341188), and Ki67 (1:500, Abcam Cat# ab16667, RRID:AB_302459).

Techniques: Derivative Assay, Control, Staining

Journal: Journal of Traditional Chinese Medical Sciences

Article Title: Exploring the molecular mechanism of Epimedium brevicornu Maxim. in treating breast cancer via network pharmacology and in vitro experiments

doi: 10.1016/j.jtcms.2024.03.005

Figure Lengend Snippet: Fig. 12. Effects of drug treatment on the expression of (A) Bax and PCNA, and (B) TBK1/NAK via western blot analysis. MDA-MB-231 cells were treated with Anhy or Iso for 24 h at the indicated concentration (20 mmol/L). Notes: Con: Control; DMSO: dimethylsulfoxide; Iso: isoliquiritigenin; Anhy: b-anhydroicaritin. Con: MDA-MB-231 cells in complete culture medium containing 1% fetal bovine serum. DMSO was used for the dissolution of Anhy and Iso. The DMSO group was included in the analysis to check for solvent interference in the results. To ensure equal loading of protein, b-actin was used as a loading control. The respective bar graphs are presented as densitometry analysis as mean (standard deviation) of experiments. *P < .05 and **P < .01 vs. the Con group.

Article Snippet: The primary antibodies: TBK1 (1:2000, Cat# 3013S), Bax (1:1000, Cat# 5023T), and PCNA (1:1000, Cat# 2586T) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Expressing, Western Blot, Concentration Assay, Control, Dissolution, Solvent, Standard Deviation