Signaling pathway of insulin-like growth factor-II as a target of molecular therapy for hepatoblastoma

Figure 1 IGF-II stimulated proliferation of hepatoblastoma cells. IGF-II was added to the medium without serum, followed by MTS assay, a modified method of MTT assay (^aP < 0.05).

Figure 2 Western blot analysis clearly shows specific bands to IGF-IR. Protein was isolated 72 h after stimulation with IGF-II (200 μg/L). The same membrane was reprobed with anti-Tubulin-α antibody to confirm an equal amount of protein loadings.

Figure 3 PPP was added to the medium 30 min prior to the stimulation with IGF-II (200 μg/L) and suppressed proliferation of Huh-6 and HepG2 (^aP < 0. 05).

Figure 4 LY294002 or Wortmannin, selective inhibitors of PI3 kinase, or PD98059, a selective inhibitor of MAP kinase, was added to the medium 30 min prior to the stimulation with IGF-II (200 μg/L). LY294002 (A) and Wortmannin (B) suppressed the proliferation of Huh-6 and HepG while PD98059 suppressed HepG2 (C)

Figure 5 Huh-6 and HepG2 dead due to apoptosis. HE staining was performed to analyze morphological changes after addition of inhibitors. Many Huh-6 cells were dead treated with PPP (60 μmol/L), LY294002 (50 μmol/L), or Wortmannin (200 μmol/L) while HepG2 with PPP, LY294002, Wortmannin, or PD98059 (20 μmol/L). Most of the dead cells had pyknotic or fragmented nuclei (arrows), suggesting apoptosis. Areas indicated by arrows were magnified (x 400).