Crocetin inhibits cell cycle G₁/S transition through suppressing cyclin D1 and elevating p27^kip1in vascular smooth muscle cells

Crocetin is a natural carotenoid compound isolated from Gardenia jasminoids Ellis. Our previous study shows that crocetin inhibits angiotensin II (Ang II)-induced vascular smooth muscle cells (VSMCs) proliferation. To further explore the mechanism by which crocetin inhibits VSMCs proliferation, in the present study we examined the effect of crocetin on cell cycle progression and cell cycle regulatory proteins. Flow cytometry analysis showed that Ang II elicited significant increase in the percentage of VSMCs in the S phase, with a concomitant decline in the percentage of VSMCs in the G(0)/G(1) phase. However, on pretreatment of VSMCs with crocetin, the percentage of VSMCs in the S phase decreased, while that in the G(0)/G(1) phase increased significantly. In addition, Ang II-induced increase of cell proliferation index was also decreased by crocetin. Western blotting analysis indicated that crocetin markedly inhibited the protein expression of cyclin D1 but not cyclin E. Crocetin also increased the level of cyclin-dependent kinase inhibitor (CDKI) p27(kip1) but not CDKI p27(waf1/cip1). In conclusion, our present results suggest that the inhibition of cell cycle G(1)/S transition in VSMCs by crocetin can be attributed, at least in part, to its suppression of cyclin D1 and elevation of CDKI p27(kip1). Copyright (C) 2009 John Wiley & Sons, Ltd.