Daphnetin has been shown to be a potent in vitro anti-proliferative agent to the human
renal cell carcinoma (RCC) cell line, A-498. In the present study, we investigated its effects on
mitogen-activated protein kinase (MAPK) signalling along with cell cycle events and cellular differentiation.
Daphnetin-activated p38, however, higher concentrations were required to inhibit ERK1/ERK2. In addition, it did not activate SAPK or induce apoptosis, but instead inhibited S phase cell cycle transition of A-498 cells at low concentrations and time of exposure. In addition, a late G(1), early S phase inhibition was observed at higher concentrations and time of exposure, indicating that the mechanism of
daphnetin-induced differentiation was concentration dependent. Increased expression of the epithelial
differentiation markers cytokeratins 8 and 18, correlated with increasing concentrations of
daphnetin, while pre-treatment with a specific p38-inhibitor, served to limit this effect. There was no evidence that
P-glycoprotein (P-gp) mediated multi-drug resistance (MDR) played a role in the anti-proliferative activity of
daphnetin. Consequently, we concluded that
p38 MAP kinase is intrinsically involved in mediating the effect of
daphnetin in A-498 cells, suggesting that this
drug may act by promotion of cellular maturation, and consequently may represent a novel low toxic approach for the treatment of poorly differentiated RCCs.