Figure 4

CD31, PCNA and TUNEL analyses for tumor tissue. (a) Tumor sections immunostained with an antibody against PCNA revealed that there were many strongly positive nuclei in control tumor tissues, whereas such nuclei were rare in tumor tissues of CPT-TMC-treated group. (b) Quantification of PCNA staining showed percentage of PCNA-positive nuclei in CPT-TMC-treated group was the lowest among the four groups (*P < 0.05, **P < 0.01). (c) Apoptosis of tumor tissues in different groups were calculated by TUNEL assays, which showed that CPT-TMC induced a significant enhancement of apoptotic cells in contrast to control therapies. (d) Quantification of TUNEL assay shows that apoptosis index of CPT-TMC-treated tumor was much higher than that of control groups (*P < 0.05, **P < 0.01). (e) Tumor sections immunostained with anti-CD31 antibody (brown) for angiogenesis assay. Representative sections were taken from tumor tissue of NS-treated, TMC-treated, CPT-treated and CPT-TMC-treated groups. (f) Histomorphometric assay for tumor microvessels revealed that MVD was significantly lower in CPT-TMC-treated group compared with the controls (*P < 0.05, **P < 0.01).