Fig. 2. Effects of low-dose Dox on tumor cell phenotype.
(A) Morphological changes induced by treatment with low-Dox. Cell and nuclear enlargement were observed after 4 days of treatment with low-dose Dox. (B) Quantitation of lipid droplet synthesis as determined by Oil-Red O staining. Cells were treated with Dox as above and analyzed by spectrophotometry. Na-butyrate represents a positive control. Absorbance is expressed as OD per million of cells. Lipid droplet synthesis was significantly enhanced by low-dose Dox. Data are represented as means ± standard deviation (n = 3, two-tailed t-test, ***p<0.0005). (C) Cell cycle analysis. Cell cycle arrest in G2 was observed following treatment with low-dose Dox. (D) Survivin expression. BT-20 cells were treated with Dox (0.1μM) for the indicated times. Cell lysates were analyzed by western blot with anti-survivin antibody. Upregulation of survivin by Dox was observed as early as 16 hrs of treatment. (E) Survivin downregulation by MN-EPPT-siBIRC5. BT-20 cells were treated with Dox, MN-siBIRC5, or MN-siSCR for 48h. The lysate proteins were analyzed for survivin and beta actin expression by western blotting. Beta-actin was used as control.