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. 2015 Sep 10;6(31):31927–31943. doi: 10.18632/oncotarget.5578

Figure 6. Icaritin inhibits tumor formation in xenografts of PLC/PRF/5 cells and primary HCC cells.

Figure 6

(A). NOD/SCID mice were implanted subcutaneously with 1×106 PLC/PRF/5 cells per mouse. After tumors reached the volume of 100 mm3, the indicated concentrations of Icaritin and the vehicle control (corn oil) were administered by gastric gavage, while Cisplatin (2 mg/kg) was administered intravenously. Tumor volume (A) and mice weights (C) were measured twice a week for 14 days after drugs administration. Points, mean (n=10); bars, SEM. **P < 0.01; ***P<0.001; Vehicle group versus other groups. (B). The tumors formed by PLC/PRF/5 cells were excised and lysed. Representative results from Western blots with the antibodies of p-Stat3 (Y705), total Stat3 and GAPDH are shown. (D). After tumors formed by PLC/PRF/5 cells reached 100mm3, Icaritin (70mg/kg) was administered by gastric gavage daily and Sorafenib (20mg/kg) was administrated intravenously daily. Tumor weights of each group were calculated 14 days after drug treatment. Columns, mean (n=5); bars, SEM. ***P<0.001. (E). Representative western blots results with the antibodies of p-Stat3 (Y705), total Stat3 and GAPDH. (F). Hep-12 cells were treated with DMSO (0) and the indicated concentrations of Icaritin for 24 h and48 h respectively and analyzed with the CCK8 assay. Each point represents mean (n=3) ± SD and data are expressed as percent of the DMSO control cells. (G). Hep-12 cells were treated with the DMSO control and Icaritin (7.5 μM) for 2 days and the 7-AAD negative cells were analyzed with EpCAM flow cytometry. The results represent two independent experiments. (H). Hep-12 cells were treated with the indicated concentrations of Icaritin for 24 h. Total cell lysates were prepared and western blots were performed using the indicated antibodies. GAPDH was used to ensure equal loading. (I). Effects of Icaritin on Hep-12 cells xenografts. NOD/SCID mice were implanted subcutaneously with Hep-12 cells (1×106/each). Icaritin (17.5 mg/kg) or the vehicle control was administered by gastric gavage daily. Tumor volume was measured twice per week for 14 days. Points, mean (n=6); bars, SEM. *, P < 0.05; Icaritin group versus vehicle group. (J). Icaritin reduces phosphorylation of Stat3 at the Tyr705 determined by IHC staining. ICT, Icaritin.