RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry.
RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry.
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DOI：
10.1152/ajpcell.00491.2004
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发表时间：
2005-08
期刊：
影响因子：
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通讯作者：
L. Liang;Á. Zsembery;E. Schwiebert
中科院分区：
文献类型：
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作者：
L. Liang;Á. Zsembery;E. Schwiebert
囊性纤维化（CF）的一种假定治疗途径是通过刺激细胞外溶液中的Ca（2+）进入激活Ca（2+）依赖性Cl（-）通道，而与CFTR功能状态无关。我们已经证明，细胞外锌和ATP诱导人气道上皮细胞胞浆Ca（2+）持续增加，转化为刺激非CF和CF人和小鼠气道上皮细胞、细胞单层和鼻粘膜的持续分泌性Cl（-）转运。基于这些研究，最可能涉及的Ca（2+）进入通道是P2 X嘌呤能受体通道。在本研究中，分子和生物化学数据显示P2 X（4）、P2 X（5）和P2 X（6）亚型在非CF（16 HBE 14 o（-））和CF（IB 3 -1）人支气管上皮细胞中共表达。其他P2 X受体Ca（2+）进入通道亚型在气道上皮、其他CF相关组织的上皮细胞模型或血管内皮中很少表达或根本不表达。在fura-2 Ca（2+）测量中，新的瞬时脂质转染介导的对P2 X（4）和P2 X（6）（但不对P2 X（5））特异的小干扰RNA片段到IB 3 -1 CF人气道上皮细胞中的递送显著抑制细胞外锌和ATP诱导的Ca（2+）内流，并且“敲低”蛋白> 65%。这些数据表明，多种P2 X受体Ca（2+）进入通道亚型在气道上皮细胞中表达。P2 X（4）和P2 X（6）可以在气道表面共组装，作为可能的治疗CF的靶点，而不依赖于CFTR基因型。
A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca(2+)-dependent Cl(-) channels via stimulation of Ca(2+) entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca(2+) in human airway epithelial cells that translates into stimulation of sustained secretory Cl(-) transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca(2+) entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X(4), P2X(5), and P2X(6) subtypes in non-CF (16HBE14o(-)) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca(2+) entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X(4) and P2X(6) (but not P2X(5)) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca(2+) entry markedly in fura-2 Ca(2+) measurements and "knocked down" protein by >65%. These data suggest that multiple P2X receptor Ca(2+) entry channel subtypes are expressed in airway epithelia. P2X(4) and P2X(6) may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype.