GNMT sensitizes HuH-7 cells to rapamycin treatment. (A, B) The effect of rapamycin on the growth of HuH-7-GFP and HuH-7-GNMT cells. Viable cells were measured by using an MTT assay. The cell viability curves were drawn by using data from d 5. The y axis represents the percentages of viable cells compared with the solvent control. **P < 0.01. Each experiment was repeated three times. (C) Effects of RAD001 on growth rates of HuH-7-GNMT and HuH-7-GFP cells in xenografts. Mean tumor volume ± standard error of the mean at a given time is shown. Treatment with RAD001 of tumors formed by inoculation of GNMT stable cells (HuH-7-GNMT + RAD001, n = 10) resulted in a significant reduction of tumor growth in comparison with the other three groups (HuH-7-GFP + RAD001, n = 8; HuH-7-GFP + placebo, n = 8; and HuH-7-GNMT + placebo, n = 5). *P < 0.05. (D) Representative photomicrographs of IHC analysis for proliferation marker Ki-67 in xenografts from (C) and quantification of Ki-67+ cells in different groups. Data are expressed as means ± SD. Means not sharing the same lowercase letters (a and c) are significantly different (P < 0.05) (bar = 100 μm). Panel A: ---○---, HuH-7-GFP + 0 (nmol/L of rapamycin);
, HuH-7-GFP + 4;
, HuH-7-GFP + 20;
, HuH-7-GFP + 100; ●, HuH-7-GNMT + 0;
, HuH-7-GNMT + 4;
, HuH-7-GNMT + 20;
, HuH-7-GNMT + 100; panel B: —○—, HuH-7-GFP; ---●---, HuH-7-GNMT; panel C: —○—, HuH-7-GFP + placebo;
, HuH-7-GFP + RAD001;
, HuH-7-GNMT + placebo;
, HuH-7-GNMT + RAD001.