NaB inhibits cell viability and proliferation of human prostate cancer DU145 cells.

<p>(A) NaB inhibits cell viability in a time- and dose-dependent manner in DU145 cells. Cancer cells were treated with medium alone or various concentrations of NaB (1, 5, 10 mmol) for various time (12, 24, 48, 72, 96 and 120 h). (B) Effect of NaB on DU145 cell proliferation. Cancer cells were treated with medium alone or various concentrations of NaB (1, 5, 10 mmol) for 48 h. (C) NaB inhibits cell viability in PC3 cells. Cancer cells were treated with medium alone or various concentrations of NaB (1, 5, 10 mmol) for various time (12, 24, 48, 72, 96 and 120 h). (D) Effect of NaB on PC3 cell proliferation. Cancer cells were treated with medium alone or various concentrations of NaB (1, 5, 10 mmol) for 48 h. (E) Effect of NaB in combination with DOC on the cell viability of DU145 and PC3 cells. Cancer cells were treated with medium alone or NaB (5 mmol) in combination with DOC (1 µmol) for 48 h. (F) Effect of NaB in combination with DOC on DU145 and PC3 cell proliferation. Cancer cells were treated with medium alone or NaB (5 mmol) in combination with DOC (1 µmol) for 48 h. Cell viability was determined by Trypan Blue exclusion assay. Cell proliferation was detected by MTT assay. NaB significantly inhibited the growth of DU145 and PC3 cells. All experiments were repeated at least three times. Data are means ± SEM. (n = 3) (Dunnett’s test, *P<0.05 <i>vs.</i> control).</p

NaB treatment induces apoptosis of DU145 cells in a dose-dependent manner.

<p>(A) Flow cytometry analysis of NaB treated DU145 cells stained with annexin V and PI. (B) Nuclear morphological observation of NaB treated DU145 cancer cells. Hoechst 33258 and a fluorescence microscope were used to observe the nuclear morphology of DU145 cells after treatment with NaB (400×). (C) Quantitative analyses of the rate of apoptosis cell. (D) NaB modulates expression of Bcl-xl and Bak in DU145 cells. (E) NaB modulates expression of Bcl-2 and Bax in DU145 cells. The expression of Bcl-xl, Bcl-2, Bax and Bak were determined by western blot analysis. All experiments were repeated at least three times. Data are means ± SEM. (n = 3) (Dunnett’s test, *P<0.05 <i>vs.</i> control).</p

NaB treatment up-regulates the expression of ANXA1 in DU145 cells.

<p>(A) ANXA1 mRNA expression is detected by qRT-PCR. NaB treatment groups significantly higher than control group (treatment by medium only). (B) Western blot analysis of ANXA1 expression in DU145 cells after NaB treatment. Data are normalized with β-actin values and are expressed as fold changes of β-actin in NaB treated group relative to control. All experiments were repeated at least three times. Data are means ± SEM. (n = 3) (Dunnett’s test, *P<0.05 <i>vs.</i> control).</p

The effect of NaB treatment on siANXA1-transfected cells.

<p>(A) Western blot analysis of ANXA1 expression in siANXA1 and siMock cells after NaB treatment. Data were normalized with β-actin values and are expressed as fold changes of β-actin in NaB treated group relative to control. (B) The proliferation of siANXA1 and siMock cells after NaB treatment. (C) Western blot analysis of the expression of Bax and Bcl-2 in siANXA1 and siMock cells. Bcl-2 and Bax expression levels were normalized to β-actin and are presented as the Bax/Bcl-2 ratio. All experiments were repeated at least three times. Data are means ± SEM. (n = 3) (Dunnett’s test, *P<0.05 <i>vs.</i> control).</p