Figure 4. CCI-779 and HCQ in combination inhibit UACC903 tumor xenograft growth.

(A) Nude mice bearing 100 mm³ tumors were given daily s.c. injection of 65 mg/kg HCQ and 0.5 mg/kg CCI-779 at the days indicated. Tumor size was measured once every three days. Each data point represents the mean ± SE tumor volume from 5 mice, 10 tumors, with tumor suppression greatest in mice treated with both CCI-779 and HCQ (*P = 0.001, **P = 0.014). (B) Representative pictures of immunohistochemical analysis showed that Ki67 was reduced in tumors treated with CCI-779, HCQ either alone or in combination. The proliferation index was calculated as the proportion of Ki67 positive cells among 200 cells in 4 randomly selected fields. Error bars represent standard errors. (*) P = 0.004; (**) P = 0.0013 (t-test) compared with the vehicle control. Active-caspase-3 staining was increased in tumors treated with HCQ, and was further increased in tumors treated with CCI-779 and HCQ. The apoptotic index was calculated as the proportion of active caspase 3 positive cells among 200 cells in 4 randomly selected fields. (*) P = 0.007; (**) P = 0.0014 (t-test) compared with vehicle control.