KN-93, a specific inhibitor of CaMKII inhibits human hepatic stellate cell proliferation in vitro

Abstract

Aim: To investigate the effects of KN-93, a CaMKII selective inhibitor on cell proliferation and the expression of p53 or p21 protein in human hepatic stellate cells.

Methods: Human hepatic stellate cells (LX-2) were incubated with various concentrations (0-50 micromol/L) of KN-93 or its inactive derivative, KN-92. Cell proliferation was measured by CCK-8 assay, and the expression of two cell cycle regulators, p53 and p21, was determined by SDS-PAGE and Western blotting.

Results: KN-93 (5-50 micromol/L) decreased the proliferation of human hepatic stellate cells in a dose-dependent manner from 81.76% (81.76% +/- 2.58% vs 96.63% +/- 2.69%, P < 0.05) to 27.15% (27.15% +/- 2.86% vs 96.59% +/- 2.44%, P < 0.01) after 24 h treatment. Incubation of 10 micromol/L KN-93 induced the cell growth reduction in a time-dependent manner from 78.27% at 8 h to 11.48% at 48 h. However, KN-92, an inactive derivative of KN-93, did not inhibit cell proliferation effectively. Moreover, analysis of cell cycle regulator expression revealed that KN-93 rather than KN-92 reduced the expression of p53 and p21.

Conclusion: KN-93 has potent inhibitory effect on proliferation of LX-2 cells by modulating the expression of two special cell cycle regulators, p53 and p21.

Figure 1

WST-8 assay showing the effect of KN-93 and KN-92 on the growth of LX-2 cells after incubation for 24 h (A) and at the indicated time (B). Data are expressed as mean ± SE of three experiments.

Figure 2

Effects of KN-93 (A) and KN-92 (B) on expression of p53 in LX-2 cells.

Figure 3

Effects of KN-93 (A) and KN-92 (B) on expression of p21 in LX-2 cells.