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1 Department of Medicine, University of California, San Diego, La Jolla, California, USA
* To whom correspondence should be addressed. E-mail: xiyuan{at}ucsd.edu.
Extracellular ATP and intracellular cAMP response element-binding protein (CREB, a transcription factor) promote cell proliferation in many cell types. The canonical-type transient receptor potential (TRPC) channels, putatively participating in forming store- and receptor-operated Ca2+ channels, have been implicated in the pulmonary vascular remodeling processes. A link among extracellular ATP, CREB activation, and TRPC4 channel expression and activity has not been shown in human pulmonary artery smooth muscle cells (PASMC). Long-term (24-48 h) treatment of human PASMC with a low dose (100 µM) of ATP, which did not trigger a transient rise in free cytosolic Ca2+ concentration ([Ca2+]cyt) when applied acutely to the cells, caused marked increases in CREB phosphorylation and TRPC4 protein expression. The time course indicated that the ATP-mediated CREB phosphorylation preceded TRPC4 upregulation, whereas transfection of a non-phosphorylatable CREB mutant abolished ATP-mediated TRPC4 expression. Furthermore, treatment of human PASMC with ATP also enhanced the amplitude of capacitative Ca2+ entry (CCE) induced by passive store depletion, whereas the siRNA specifically targeting on TRPC4 attenuated ATP-mediated increases in TRPC4 expression and CCE amplitude, and inhibited ATP-induced PASMC proliferation. These data suggest that low-dose ATP exerts part of its mitogenic effect in human PASMC via CREB-mediated upregulation of TRPC4 channel expression and activity, and the subsequent increase in CCE and [Ca2+]cyt.
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