Potentiation effect of Bcl-2-ASO with TAX and DEX.

Bcl-2-ASO potentiates TAX- and DEX-induced apoptosis (A) and down-regulation of Bcl-2 (B) in ARH-77 myeloma cells. ARH-77 cells were cultured for 3 days with or without 5 μg/mL Bcl-2-ASO (AS) or reverse sense oligodeoxynucleotides (RS). Cultures were continued for additional 2 days with or without 0.1 μM TAX or 0.5 μM DEX. Cells were then harvested and analyzed for apoptosis and for Bcl-2 protein expression. Cultures treated for 1 day or 2 days with TAX or DEX alone were also prepared and analyzed for apoptosis and for Bcl-2 expression. Bcl-2-ASO was replenished every other day. Bcl-2 protein was identified as a 26 kDa migrating band. Note that in TAX-treated cells a slow migrating band of about 28 kDa is visible. This was identified as the widely documented phosphorylated Bcl-2 12 and was included in the scanning for Bcl-2 content. Note the additive effect of drugs and Bcl-2-ASO on increased apoptosis (A) and decreased Bcl-2 protein (B). Other experimental details are as in Figure 1. Error bars represent ± SD for at least 3 experiments.