Figure 2

Statins inhibited the expressions of MCM7 and RB and induced apoptosis and DNA damage. (a) A small molecular drug screening was carried out and SVA showed strongest inhibition effect on MCM7 protein. (b) MCM7 and RB expressions were inhibited after Hep3B cells were treated for 24 or 72 h with 5 or 10 μM Simvastatin, 20 and 40 μM Atorvastatin, 50 and 100 μM Fluvastatin, 10 and 20 μM Lovastatin, and 10 and 20 μM Pravastatin. (c) Hep3B cells were treated with Simvastatin (10 μM), Atorvastatin (40 μM) and Pravastatin (20 μM) for 72 h, harvested and incubated with PI, and then analyzed by FACS. Statistical analysis showed differences in the G0/G1 phase and the sub-diploid peak between drug-treated cells and control cells. (d) Hep3B cells were treated with Simvastatin (5 or 10 μM) or Atorvastatin (20 or 40 μM) for 24 or 72 h, and cell extracts were analyzed by immunoblotting. The expression of MCM7, RB, p-RB, cyclin D1, PCNA and CDC45 were reduced and the expressions of p21 and p27 were increased. (e) Hep3B cells were treated as above; cell extracts were analyzed by immunoblotting with Bcl2, γ-H2AX, PARP or cleaved-caspase3 antibodies. (f) Hep3B cells were treated with 10 μM Simvastatin or 40 μM Atorvastatin for 72 h, then incubated with γ-H2AX antibody and imaged by laser confocal microscopy (scale bars, 10 μm)