Figure 6.

Effect of combination of Mithramycin and bicalutamide on cell growth and expression of AR, PSA, and p21. Panel A: Synergistic combination of Mithramycin and bicalutamide by MTT. Exponentially growing AD and AI cells in 96-well dishes were were exposed for three days to serial dilution of Mithramycin or bicalutamide, or Mithramycin plus bicalutamide at ratio of 1:3300. The cell growth was measured by MTT, and the percentages of inhibition (1-T/C%) were calculated, and combination index (CI) analyzed by a PC program, CalcuSyn, of Biosoft (see ligand for Figure 5). Panel B: Effects of the combination of mithramycin with bicalutamide on expressions of AR, PSA, and p21. AD and AI cells at exponential growth were exposed to indicated concentration of Mithramycin, bicalutamide alone or their combination for 24 hrs. The cells were harvested, washed, and total proteins extracted. Fifty μg were subjected to SDS-PAGE, electro-transferred to nitrocellulose membrane, and immuno-blotted with antibodies against AR, PSA, and p21 (Santa Cruz and DAKO).