Figure 9.

Inactivation of SKP2 or non-degradable p27^KIP1 is detrimental to the growth of HCC cells in vitro. (A) Transfection of SKP2dn triggers the upregulation of p27^KIP1 levels in SNU449 cells, as detected by Western blot analysis. As expected, transient transfection of SKP2dn resulted in the expression of a truncated form of SKP2 (transfected) with a lower molecular weight than the endogenous protein. β-Actin was used as a loading control. Transfection of SKP2dn reduces proliferation (B) and increases apoptosis (C) in the same cells. (D) Transfection of p27^KIP1−187A results in the appearance of a second band (transfected) with a higher molecular weight than the endogenous p27^KIP1 protein in the SNU449 cell line. β-Actin was used as a loading control. Similar to that described for SKP2dn, transfection of p27^KIP1−187A decreases proliferation (E) and augments apoptosis (F) in the same cell line. Student’s t-test: p < 0.0001 *** vs. empty vector (control). Experiments were conducted three times in triplicate.