Figure 4

Treatment with P4HA inhibitor attenuates breast cancer cell proliferation and invasiveness. (A) Phase images of control and 1,4-DPCA treated ZR-75-1 (20 μM 1,4-DPCA), T4-2 (10 μM 1,4-DPCA), MDA-MB-157 (20 μM 1,4-DPCA) and MDA-MB-231 (10 μM 1,4-DPCA) cells in 3D culture. The cells treated with 1,4-DPCA formed smaller and less aggressive structures compared to control cells (scale bar, 50 μm). (B) Quantification of colony size of control and 1,4-DPCA treated ZR-75-1 and T4-2 cells in 3D culture by measuring the diameter of at least 50 colonies. Treatment with 1,4-DPCA reduced the colony size. (C) Ratio of polarized colonies in control and 1,4-DPCA treated T4-2 cells in 3D culture. (D) Quantification of the invaded branches of control and the 1,4-DPCA treated MDA-MB-157 and MDA-MB-231 cells in 3D culture by measuring the branches in at least 50 colonies. Cells treated with 1,4-DPCA had decreased invasive branch number. (E) EdU-staining was used to analyze the proliferation of control and the 1,4-DPCA treated ZR-75-1, T4-2, MDA-MB-157 and MDA-MB-231 cells in 3D culture. Treatment with 1,4-DPCA decreased proliferation of these four cell lines (*p < 0.05; **p < 0.01).