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首页> 外文期刊>Molecular biology of the cell >Extracellular ATP causes ROCK I-dependent bleb formation in P2X(7)-transfected HEK293 cells
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Extracellular ATP causes ROCK I-dependent bleb formation in P2X(7)-transfected HEK293 cells

机译：细胞外ATP在P2X（7）转染的HEK293细胞中引起ROCK I依赖的气泡形成

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摘要

The P2X(7) ATP receptor mediates the cytotoxic effect of extracellular ATP. P2X(7)-dependent cell death is heralded by dramatic plasma membrane bleb formation. Membrane blebbing is a complex phenomenon involving as yet poorly characterized intracellular pathways. We have investigated the effect of extracellular ATP on HEK293 cells transfected with the cytotoxic/pore-forming P2X(7) receptor. Addition of ATP to P2X(7)-transfected, but not to wt P2X(7)-less, HEK293 cells caused massive membrane blebbing within 1-2 min. UTP, a nucleotide incapable of activating P2X(7), had no early effects on cell shape and bleb formation. Bleb formation triggered by ATP was reversible and required extracellular Ca2+ and an intact cytoskeleton. Furthermore, it was completely prevented by preincubation with the P2X(7) blocker oxidized ATP. It was recently observed that the ROCK protein is a key determinant of bleb formation. Preincubation of HEK293-P2X(7) cells with the ROCK blocker Y-27632 completely prevented P2X(7)-dependent blebbing. Although ATP triggered cleavage of the ROCK I isoform in P2X(7)- transfected HEK293 cells, the wide range caspase inhibitor z-VAD-fluoromethylketone had no effect. These observations suggest that P2X(7)-dependent plasma membrane blebbing depends on the activation of the serine/threonine kinase ROCK I. [References: 45]
机译：P2X（7）ATP受体介导细胞外ATP的细胞毒性作用。 P2X（7）依赖的细胞死亡是由戏剧性的质膜气泡形成预示的。膜起泡是一种复杂的现象，涉及尚未充分表征的细胞内途径。我们已经研究了细胞外ATP对HEK293细胞转染细胞毒性/形成孔的P2X（7）受体的影响。 ATP添加到P2X（7）-转染，但不添加到wt P2X（7）-少，HEK293细胞在1-2分钟内引起大量膜起泡。 UTP是一种不能激活P2X（7）的核苷酸，对细胞形状和气泡形成没有早期影响。由ATP触发的小脑形成是可逆的，需要细胞外Ca2 +和完整的细胞骨架。此外，通过与P2X（7）阻断剂氧化的ATP预先孵育，可以完全避免这种情况。最近观察到，ROCK蛋白是起泡形成的关键决定因素。与ROCK阻滞剂Y-27632一起预孵育HEK293-P2X（7）细胞可完全防止P2X（7）依赖性起泡。尽管ATP触发了P2X（7）转染的HEK293细胞中ROCK I亚型的切割，但宽范围的半胱天冬酶抑制剂z-VAD-氟甲基酮没有作用。这些观察结果表明依赖P2X（7）的质膜起泡取决于丝氨酸/苏氨酸激酶ROCK I的激活。[参考文献：45]

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