Fig. 5
In vivo effects of nanomedicines on proliferation activities in HCC liver orthotropic and subcutaneous models. a Tumor weight measured at day 20 after treatment began in Hep3B, SMMC-7721 and BEL7402 liver orthotropic model. b Protein expression levels of PBOV1 determined by western blotting assay in Hep3B, SMMC-7721, and BEL7402 liver orthotropic tumor tissues from mice after treatment for 20 days. c Kinetic of the liver orthotropic tumor growth measured by MRI T2 scan at 2 h after nanomedicine injection (nanomedicines injected once every day from day 11; to visulaize the tumors of PBS group by MRI, TOV nanomedicine was injected 2 h before MRI scan). d First panel, MRI T2 scan for liver orthotropic tumors; second panel, optical imaging of liver showing orthotropic tumors at day 20 after treatment began. e Protein expression levels of proliferation marker survivin, cell cycle marker cyclin D1 and apoptotic marker cleaved-caspase 3 in Hep3B liver orthotropic tumor tissues at day 20 after treatment began. f First and second panel from top down, protein expressions of PBOV1 determined by IHC in liver orthotropic tumors in various treatment groups, and magnified view showing location of PBOV1 expression inside cancer cells (Scale bar, 50 μm); third and fourth panel from top down, in vivo tumor cell proliferation abilities measured by immunofluorescence staining assays for Bromodeoxyuridine (BrdU) and Ki67; fifth panel from top down, tumor apoptosis detected by TUNEL assay (Scale bar, 20 μm) (Blue, DAPI-indicating nuclei; red, Cy5 indicating BrdU location; green, FITC indicating Ki67 protien or apoptotic nuclei). g Observation of the tumor inhibition effect in subcutaneous tumor model during the 20 days of treatment (see also Supplementary Fig. 6). Data were represented as means ± S.D. Error bars represent the S.D. values (n = 12; *P < 0.05; ns, not significant. Significant differences were analyzed by two-way ANOVA with multiple comparisons for c and g; Student’s t test for (a, c and g), compared with PBS control group)
