FIGURE 6 from Identification of an Imidazopyridine-based Compound as an Oral Selective Estrogen Receptor Degrader for Breast Cancer Therapy
Year: 2023
Type: other
Abstract: X15695 and fulvestrant downregulate ERα level and proliferation of breast cancer cell proliferation A, Western blot analysis of ERα in extracts of MCF-7 cells treated for the indicated timepoints with 1 μmol/L of fulvestrant or X15695 or DMSO (vehicle). β-actin was used for the loading control. The protein signals were quantified and presented in percentages as the signal of ERα relative to the β-actin signal (right). The value at zero time of treatment was set to 100%. The values are the mean ± SEM (n = 3; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001). B, Western blot analysis of ERα in extracts of MCF-7 cells treated with the indicated concentration of fulvestrant or X15695 for 8 hours. β-actin was used for the loading control. The protein signals were quantified and presented in percentages relative to the vehicle-treated sample (right). The values are the mean ± SEM (n = 3; ****, P ≤ 0.0001). C, Measurement of cell viability by MTT assay of tamoxifen resistant cells (TRMCF-7) treated with the indicated concentrations of X15695, fulvestrant, and tamoxifen for 48 hours. Five replicates were used for each experiment, and the experiment was repeated three times (**, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; ns, nonsignificant). D, Representative image of a clonogenic assay showing the effect of the indicated concentrations of X15695, fulvestrant, and tamoxifen on clonal expansion of tamoxifen-resistant MCF-7 cells. E, Quantification of colonies in the clonogenic assay described in D. F, Representative images of some of the tumors obtained in a mouse MCF-7 tumor xenograft after treatment with vehicle and X15695. G, Tumor volumes of established subcutaneous MCF-7 xenografts treated daily via oral gavage with vehicle or X15695 (30 mg/kg body weight). The values are the mean ± SEM (*, P = 0.05; n = 14). H, Tumor weights at the end of treatment were recorded and presented in the bar chart. The values are the means ± SEM (*, P = 0.05; n = 14). I, Mouse body weights measured twice weekly for all animals over 16 days. Presented are the mouse body weight changes, in percentages, for each group. J, Western blots analysis of ERα and p53 from the extracts of select tumors at the end of treatment. Anti-vinculin antibody was used to determine the level of protein loaded. Quantification of ERα (K) and p53 (L) protein levels in select tumors relative to the vinculin loading control. The results are the mean ± SEM. *, P ≤ 0.05; ***, P ≤ 0.001; n = 12.
Authors: Mengwu Pan, Valeria Solozobova, Nane C. Kuznik, Nicole Jung, Simone Gräßle, Victor Gourain, Yvonne M. Heneka, Christina A. Cramer von Clausbruch, Olaf Fuhr, Ravi S. N. Munuganti, Danilo Maddalo, Christine Blattner, Antje Neeb, Adam Sharp, Laura Cato, Carsten Weiss, Rinath M. Jeselsohn, Veronique Orian-Rousseau, Stefan Bräse, Andrew C. B. Cato
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4385303140