Research Paper Volume 13, Issue 9 pp 12479—12492

Figure 6. SUMO1P3 depletion impeded HCC growth and metastasis in vivo. Four-week-old male SCID mice were inoculated subcutaneously into the hind flanks or injected via tail vein with MHCC97H-luc cells with stably expressing shNC or shSUMO1P3-2. (A) Tumor volumes were monitored and calculated every week for 6 weeks. (B) Representative images of the mice (Mock and pENTR (n=10), or pENTR-shSUMO1P3 and pENTR-shSUMO1P3 (n=10)) photographed with an IVIS Imaging System at days 14 after inoculation. (C) Representative general photographs and HE staining assay of the harvested primary tumors at 6 weeks after subcutaneous xenografting. 100 × magnification in HE sections. (D) TUNEL assay was carried out to determine cell apoptosis in the tumor tissues of the xenografts. The percentage of TUNEL-positive cells was calculated. (E, F) At 8 weeks after injection through the tail vein, the lungs were harvested and photographed, and the numbers of pulmonary metastatic nodules was counted. (G) SUMO1P3 expression in tumor tissues was measured via qPCR assay. GAPDH was used as the endogenous control. (H) Representative results of Western blot analyses of cyclin D1, p-Akt, Akt, Bax, Bcl-2, cl-caspase-3, caspase-3, E-cadherin, vimentin, MMP-2, and MMP-9 in tumor tissues. β-actin was used as endogenous control. All data are represented as the mean ± SD of three replicates. *P < 0.05 vs. Mock or pENTR group.