Figure 5.

Titration Experiments Using the AR-ASO and 6BIO Demonstrate a Reduced AR Expression in Prostate Cancer Cells

Western analyses of lysates harvested from LNCaP (A–C) or LAPC-4 cells (D). (A) LNCaP cells treated with (+) or without (−) 1 μM 6BIO or 2 μM AR-ASO. The combined treatment of 6BIO and the AR-ASO reduced AR protein expression by approximately 70%. Either 6BIO or AR-ASO alone reduced the AR protein expression by only approximately 25%. Values of protein reduction were calculated with ImageJ and normalized to the GAPDH as the loading control and to the control, untreated cells. (B) LNCaP cells treated with 1 μM 6BIO with (+) or without (−) the indicated concentrations of AR-ASO. (C) LNCaP cells treated with 1 μM AR-ASO with (+) or without (−) the indicated concentrations of 6BIO. (D) LAPC-4 cells with 5 μM AR-ASO with (+) or without (−) the indicated concentrations of 6BIO. In all experiments, the AR-ASO was delivered by gymnosis for 2 days. Experiments were repeated a minimum of two times with consistent results. (E) Combination index (CI) plot calculated with the CI equation algorithms using CompuSyn software.³⁵ The values were derived from the quantitation of AR protein suppression by the combined use of 6BIO and AR-ASO (Figure S2). CI = 1 indicates an additive effect of the drugs; CI > 1 indicates an antagonistic effect; CI < 1 demonstrates synergism.