Fig. 7.

Efficacy of BC-LI-0186 to rapamycin-resistant cancer cells. a HCT116 MW (mTOR WT) and MM (mTOR S2035I) cells were treated with rapamycin, BC-LI-0186, and INK128 at the indicated concentrations (nM) for 6 h. The cell lysates were subjected to immunoblotting analysis with anti-mTOR, anti-p-S6K, anti-S6K, and anti-actin antibodies. b The level of p-S6K in HCT MW cells shown in a was quantified and displayed as graph. c The level of p-S6K in HCT MM cells shown in a was quantified and displayed as graph. d Cell growth GI₅₀ and cell death EC₅₀ values of BC-LI-0186 and rapamycin against HCT116 MW and MM cells. e GTP-agarose bead pull-down assays were used to monitor the GTP-bound RagD in HCT116 MW and MM cells. After cells were treated with 10 μM BC-LI-0186, 100 nM rapamycin, or 10 μM INK128, the cell lysates were incubated with GTP-agarose beads and the proteins precipitated with the beads were analyzed by immunoblotting with anti-RagD or anti-ARF1 antibodies. f HCT116 MM cells were injected subcutaneously to nude mice. BC-LI-0186 and rapamycin were intraperitoneally administered into the mice at the indicated doses every day (n/group = 6). Tumor volume was measured every other day for 2 weeks. The error bars represent mean ± S.D. *p < 0.05; **p < 0.01; ***p < 0.001 (vs. vehicle)