Figure 8.

Rescued erbB-2 Abrogates Oncogenic Action of miR372

(A) Western blotting analysis with anti-erbB-2, anti-CDK2, and anti-DNMT1 in liver cancer cell line Hep3B cells infected with rLV, rLV-miR372, and rLV-miR372 plus pGFP-V-RS-erbB-2, respectively. β-actin as internal control is shown. (B) coIP with anti-CDK2 followed by western blotting with anti-cyclin E in the Hep3B cells infected with rLV, rLV-miR372, and rLV-miR372 plus pGFP-V-RS-erbB-2, respectively, is shown. IgG IP as negative control is shown. INPUT refers to western blotting with anti-CDK2. (C) Proliferation assay was performed using the CCK8. Data are means of value from three independent experiments; mean ± SEM. **p < 0.01; *p < 0.05. (D) Cell BrdU assay is shown. Data are means of value from three independent experiments, mean ± SEM. **p < 0.01; *p < 0.05. (E) (Upper) The photography of colonies from the cell lines indicated in left is shown. (Lower) Cell plate colony formation ability assay is shown. Data are means of value from three independent experiments, mean ± SEM. **p < 0.01; *p < 0.05. (F) The xenograft tumors weight (gram) in the three groups is shown. Data were means of value from six BALB/c mice; mean ± SEM; n = 6; *p < 0.05; **p < 0.01. (G) The xenograft tumors appearance time (days) in the three groups is shown. Data were means of value from six BALB/c mice; mean ± SEM; n = 6; *p < 0.05; **p < 0.01. (H) (a) A portion of each xenograft tumor was fixed in 4% formaldehyde and embedded in paraffin, and the micrometers of sections (4 μm) were made for H&E staining and PCNA staining (original magnification ×100). (b) PCNA and Ki67 positive rate is shown.