Am J Physiol Cell Physiol AJP: Regulatory, Integrative and Comparative Physiology
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Am J Physiol Cell Physiol 281: C157-C165, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 1, C157-C165, July 2001

Bcl-2 decreases voltage-gated K+ channel activity and enhances survival in vascular smooth muscle cells

Daryoush Ekhterae*, Oleksandr Platoshyn*, Stefanie Krick*, Ying Yu*, Sharon S. McDaniel, and Jason X.-J. Yuan

Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of California School of Medicine, San Diego, California 92103

Cell shrinkage is an incipient hallmark of apoptosis in a variety of cell types. The apoptotic volume decrease has been demonstrated to attribute, in part, to K+ efflux; blockade of plasmalemmal K+ channels inhibits the apoptotic volume decrease and attenuates apoptosis. Using combined approaches of gene transfection, single-cell PCR, patch clamp, and fluorescence microscopy, we examined whether overexpression of Bcl-2, an anti-apoptotic oncoprotein, inhibits apoptosis in pulmonary artery smooth muscle cells (PASMC) by diminishing the activity of voltage-gated K+ (Kv) channels. A human bcl-2 gene was infected into primary cultured rat PASMC using an adenoviral vector. Overexpression of Bcl-2 significantly decreased the amplitude and current density of Kv currents (IKv). In contrast, the apoptosis inducer staurosporine (ST) enhanced IKv. In bcl-2-infected cells, however, the ST-induced increase in IKv was completely abolished, and the ST-induced apoptosis was significantly inhibited compared with cells infected with an empty adenovirus (-bcl-2). Blockade of Kv channels in control cells (-bcl-2) by 4-aminopyridine also inhibited the ST-induced increase in IKv and apoptosis. Furthermore, overexpression of Bcl-2 accelerated the inactivation of IKv and downregulated the mRNA expression of the pore-forming Kv channel alpha -subunits (Kv1.1, Kv1.5, and Kv2.1). These results suggest that inhibition of Kv channel activity may serve as an additional mechanism involved in the Bcl-2-mediated anti-apoptotic effect on vascular smooth muscle cells.

apoptotic volume decrease; pulmonary artery smooth muscle cells; current density of voltage-gated potassium channels

* D. Ekhterae, O. Platoshyn, S. Krick, and Y. Yu contributed equally to this work.




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