Fig. 2. Rapamycin and perifosine combination overcomes the growth and survival advantage in MM.1S cells conferred by IL-6 and IGF or BMSCs.
(A) MM.1S cells were cultured for 6 hours in control media (C), rapamycin 10 nM (R), perifosine 5 uM (P), or rapamycin and perifosine combination (R+P). Cells were then stimulated with IL-6 (10 ng/ml) or IGF-1 (25 ng/ml) for 10 min. Whole-cell lysates were subjected to Western blotting using anti-p-Akt and anti-Akt antibodies.
(B) MM.1S cells were cultured for 48 hours in control media, or rapamycin 10 nM, perifosine 5 uM, or their combination in the presence or absence of IL-6 (10 ng/ml) or IGF-1 (25 ng/ml). Cell proliferation was assessed by [3H]-thymidine uptake; data represent means (+/-SD) of triplicate cultures. Combination indices (CIs) were calculated based upon the isobologram generated using the Chou-Talalay method.
(C) MM.1S cells were cultured in control media (C), rapamycin 10 nM (R), perifosine 5 uM (P), rapamycin and perifosine combination (R+P) in the presence or absence of BMSCs. MM.1S cells were harvested, lysed, and subjected to Western blotting using anti-p-Akt and anti-Akt antibodies.
(D) MM.1S cells were cultured for 48 hours in control media, rapamycin 10 nM, perifosine 5 uM, or their combination in the presence or absence of BMSCs. Cell growth was assessed by [3H]-thymidine uptake; data represent means (+/-SD) of triplicate cultures. Combination indices (CIs) were calculated based upon the isobologram generated using the Chou-Talalay method.