Mitogenic modulation of Ca2+ -activated K+ channels in proliferating A7r5 vascular smooth muscle cells

1 Modulation of Ca2+-activated K+ channels (KCa) has been implicated in the control of proliferation in vascular smooth muscle cells (VSMC) and other cell types. In the present study, we investigated the underlying signal transduction mechanisms leading to mitogen-induced alterations in the expression pattern of intermediate-conductance KCa in VSMC. 2 Regulation of expression of IKCa/rKCa3.1 and BKCa/rKCa1.1 in A7r5 cells, a cell line derived from rat aortic VSMC, was investigated by patch-clamp technique, quantitative RT–PCR, immunoblotting procedures, and siRNA strategy. 3 PDGF stimulation for 2 and 48 h induced an 11- and 3.5-fold increase in rKCa3.1 transcript levels resulting in a four- and seven-fold increase in IKCa currents after 4 and 48 h, respectively. Upregulation of rKCa3.1 transcript levels and channel function required phosphorylation of extracellular signal-regulated kinases (ERK1/2) and Ca2+ mobilization, but not activation of p38-MAP kinase, c-Jun NH(2)-terminal kinase, protein kinase C, calcium-calmodulin kinase II and Src kinases. 4 In contrast to rKCa3.1, mRNA expression and functions of BKCa/rKCa1.1 were decreased by half following mitogenic stimulation. Downregulation of rKCa1.1 did not require ERK1/2 phosphorylation or Ca2+ mobilization. 5 In an in vitro-proliferation assay, knockdown of rKCa3.1 expression by siRNA completely abolished functional IKCa channels and mitogenesis. 6 Mitogen-induced upregulation of rKCa3.1 expression is mediated via activation of the Raf/MEK- and ERK-signaling cascade in a Ca2+-dependent manner. Upregulation of rKCa3.1 promotes VSMC proliferation and may thus represent a pharmacological target in cardiovascular disease states characterized by abnormal cell proliferation. British Journal of Pharmacology (2006) 148, 909–917. doi:10.1038/sj.bjp.0706793