Figure 5

Effects of SPRY4-IT1 on ER(−) breast cancer cell proliferation and apoptosis in vitro. (A) The relative ZNF703 expression level in breast cancer cell lines (MDA-MB-231, MDA-MB-435S and MCF-7) compared with the level in human breast epithelial cells (MCF-10A) was assessed by qRT-PCR. (B, C) An MTT assay was performed to determine the proliferation of MDA-MB-231 and MDA-MB-435S cells treated with scrambled siRNA and si-ZNF703. The data are presented as the means ± S.D. from three independent experiments. (D, E) Colony-forming growth assays were performed to determine the proliferation of MDA-MB-231 and MDA-MB-435S cells treated with scrambled siRNA and si-ZNF703. The colonies were counted and captured. (F, G, H) Proliferating MDA-MB-231 and MDA-MB-435S cells treated with scrambled siRNA or si-ZNF703 were labeled with EdU. The Click-it reaction revealed EdU staining (red). The cell nuclei were stained with Hoechst 33342 (blue). The images are representative of the results obtained. (I, J) The percentage of apoptotic cells was determined by flow cytometric analysis. The data are presented as the means ± SD from three independent experiments. (K) An MTT assay was performed to determine the proliferation of MDA-MB-231 cells treated with empty vector and pcDNA-ZNF703. (L, M) Colony-forming growth assays were performed to determine the proliferation of MDA-MB-231 cells treated with empty vector and pcDNA-ZNF703. The colonies were counted and captured. Three independent experiments were performed for each assay. *P < 0.05 and **P < 0.01.