Fig. 5From: The PRMT5-LSD1 axis confers Slug dual transcriptional activities and promotes breast cancer progressionCombination of SP2509 and EPZ015666 synergistically inhibits tumor progression of breast cancer cells (A and B) Immunoblot analysis of the indicated proteins in MDA-MB-231 (A) and SUM159 (B) cells treated with DMSO, SP2509 (1 μM), EPZ015666 (1 μM) or SP2509 plus EPZ015666 for 72 h. (C and D) Expression of the indicated genes was measured by qPCR in MDA-MB-231 (C) and SUM159 (D) cells treated with DMSO, SP2509 (1 μM), EPZ015666 (1 μM) or SP2509 plus EPZ015666 for 72 h. (E and F) MDA-MB-231 (E) and SUM159 (F) cells treated with DMSO, SP2509 (1 μM), EPZ015666 (1 μM) or SP2509 plus EPZ015666 for 72 h. The invasiveness of these cells was analyzed with a matrigel-coated chamber invasion assay. Representative photomicrographs are shown in the left. The histograms show the mean ± SD of the fold changes of numbers of invasive cells in each sample over that of control sample from three independent experiments. ***P < 0.001, as determined by Student’s t test. (G) MDA-MB-231 cells stable expressing firefly luciferase were inoculated orthotopically into the fourth mammary fat pad of 6-week-old female nude mice. The mice were treated with Vehicle, SP2509 (25 mg/kg, i.p.), EPZ015666 (100 mg/kg, i.p.) and combination of SP2509 and EZP015666 (n = 5) starting from day 10 post transplantation. The drugs were administered on days 10, 13, and 17 in an eight days repeating cycle for three cycles. Primary tumors were quantified by bioluminescence imaging on day 41 after initial implantation. Representative in vivo (upper) and in vitro (middle) tumor bioluminescent images, and excised tumors (lower) from each of group are shown. (H) The bar graphs show the mean ± SD of the primary tumor volume from mice in G. (I) The graph depicts the mean tumor growth of mice in G which received indicated treatment on the days marked by the black arrows. (J) Representative ex vivo bioluminescence imaging of the lungs removed from mice in G on day 41. (K) The bar graphs show the mean ± SD ex vivo lung photon flux of mice in G for each group. (L) MDA-MB-231 cells stable expressing firefly luciferase were intravenously injected into female nude mice. The mice were treated as described in G. Lung metastasis was quantified using bioluminescence imaging on day 41 after initial implantation. Representative in vivo (upper) and ex vivo lungs (lower) bioluminescence imaging from each of group are shown. (M) The bar graphs indicate the mean ± SD ex vivo lung photon flux of mice in L for each group. For H, I, K and M, *p < 0.05, ***P < 0.001, Dunnett’s multiple comparison testBack to article page