Els of putative target proteins in livers from AKTcMET mice. (Magnifications: 100and 200 Scale bar: 100 ). Abbreviations: H E, hematoxylin and eosin staining; Pre, pretreatment.Tumor growth was monitored in AKTcMET mice till 4 weeks just after injection, when the mice show a moderate tumor burden (average liver weight 4 g) (Figure 1A,B). Subsequently, AKTcMETCancers 2019, 11, x4 of(Magnifications: 100and 200 Scale bar: one hundred m). Abbreviations: H E, hematoxylin and eosin staining; Pre, pretreatment.Cancers 2019, 11, 930 4 ofTumor growth was monitored in AKTcMET mice until 4 weeks right after injection, when the mice display a moderate tumor burden (average liver weight four g) (Figure 1A,B). Subsequently, AKTcmice had been randomly separated into three cohorts. A group of mice at 4at four weeks postinjection MET mice have been randomly separated into three cohorts. A group of mice weeks postinjection was harvested as a `pretreatment’ cohort, whilewhile the remaining two Bcma Inhibitors medchemexpress groups have been continually treated was harvested as a `pretreatment’ cohort, the remaining two groups were continually treated with either vehiclevehicle or sorafenib for three weeks (Figure 1A). Interestingly,we located that tumor continued with either or sorafenib for 3 weeks (Figure 1A). Interestingly, we found that tumor continued to grow with sorafenib (30 mgkgday) remedy. All vehicle as also sorafenibtreated mice miceto to develop with sorafenib (30 mgkgday) treatment. All car effectively as as sorafenibtreated had had to become euthanized by three three weeks treatment due on account of higher tumortumor burden. In AKTcMET mice, euthanized by weeks of of remedy to high liver liver burden. In AKTcMET mice, tumor nodules have been diffused andand colliding, with no surrounding capsules; a consequence, it was tumor nodules had been diffused colliding, with no surrounding capsules; as as a consequence, it was not possible to accurately count surface tumor nodule number in these mice (Figure 1C, 1C, panels). not possible to accurately count the the surface tumor nodule quantity in these mice (Figureright suitable As panels). As most (more than 90 ) from the liver parenchyma was occupied by the tumor cells, we employed general most (over 90 ) in the liver parenchyma was occupied by the tumor cells, we utilized overall liver liver weight as the measure of tumor burden. This system has been shown to accurately reflect HCC weight as the measure of tumor burden. This process has been shown to accurately reflect HCC burden in this murine liver tumor model by independent burden in this murine liver tumor model by independentgroups [25,26]. We discovered that thethe sorafenibgroups [25,26]. We discovered that sorafenibtreated cohort had larger tumor burden than the pretreatment cohort, and similar tumor burden treated cohort had larger tumor burden than the pretreatment cohort, and equivalent tumor burden was found in sorafenib and vehicletreated mice (Figure 1B,C). At the D-Lysine monohydrochloride Purity & Documentation molecular level, sorafenib did was identified in sorafenib and vehicletreated mice (Figure 1B,C). At the molecular level, sorafenib not inhibit pERK or pAKT expression in the mouse liver tissues (Figure 1D). In the cellular level, didn’t inhibit pERK or pAKT expression in the mouse liver tissues (Figure 1D). At the cellular sorafenib treatment didn’t impact HCC cell proliferation, but was capable to induce apoptosis (Figure level, sorafenib remedy did not affect HCC cell proliferation, butsorafenibtreated mice, it was 2A,B). Nonetheless, as the cell apoptosis rate was reasonably low even in was able to induce.