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. 2018 Jan 9;9(8):7902–7917. doi: 10.18632/oncotarget.24083

Figure 1. HPA3P induces cell death in human colon cancer cells.

Figure 1

(A) All of the colon cancer cell lines were treated with different concentrations of HPA3, HPA3P, and HPA3P2 for 24 h. The effects of HPA3, HPA3P, and HPA3P2 on cell viability in the indicated colon cancer cell lines were measured by MTT assay. The data are shown as the mean ± SEM. *p < 0.05 and **p < 0.01 compared with control. (B) Cell death induction in colon cancer cell lines treated with HPA3P (LoVo and HT-29, 30 μM; SW480 and HCT116 p53+/+, 50 μM) was assessed by flow cytometry using annexin V and PI. (C) All cells were treated with the indicated concentrations of HPA3P for 24 h. All cell lines were treated with staurosporine, which served as a positive control. Whole-cell lysates were prepared, and apoptosis was assessed by western blot analysis using anti-cleaved caspase-3, anti-cleaved PARP, and GAPDH antibodies. (D) Anchorage-independent growth in the HPA3P-treated colon cancer lines was assessed by colony formation assay. Colony formation was observed 10 days after plating. Images were photographed using a camera attached to a Nikon SMZ800 stereomicroscope (magnification, 4×). (E) Statistical analysis was performed to quantify relative colony formation in the HPA3P-treated and non-treated cell lines. Colonies were counted using ImageJ software. The results represent the average number of colonies from three replicated experiments. The data are shown as the mean ± SD. **p < 0.01 compared with control.