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Demonstration of Proteolytic Activation of the Epithelial Sodium Channel ENaC by Combining Current Measurements with Detection of Cleavage Fragments
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细胞外血清蛋白酶激活了对胺不敏感的ENaC通道,并降低了人类白血病细胞系的迁移和入侵能力.

Daria V Lysikova1, Polina I Kirillova1, Irina O Vassilieva2

  • 1Institute of Cytology, Russian Academy of Sciences, 194064 Tikhoretsky Ave. 4, St. Petersburg, Russia.

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概括

细胞外血清蛋白酶如α-chymotrypsin和等离子体激活白血病细胞中的上皮质通道 (ENaCs),减少它们的迁移和入侵. 这突显了血清蛋白酶在通过ENaC道调节癌细胞运动中的新作用.

关键词:
表面上皮质Na+通道人类白血病细胞移民 移民 迁移 迁移塑塑是一种塑.这是一种α-chymotrypsin.

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科学领域:

  • 分子生物学分子生物学
  • 癌症研究 癌症研究
  • 离子通道生理学 离子通道生理学

背景情况:

  • 皮质通道 (ENaCs) 与癌细胞的增殖,亡和运动有关.
  • 之前的研究表明,青素刺激了白血病K562细胞中对氨酸不敏感的ENaC类通道.
  • 各种血清蛋白酶在调节血细胞中的通道和癌症进展中的作用尚不清楚.

研究的目的:

  • 为了研究血清蛋白酶α-chymotrypsin和等离子体对白血病细胞中的通道的影响.
  • 确定蛋白酶激活通道在白血病细胞迁移和入侵中的功能作用.
  • 阐明 δ-ENaC 参与蛋白酶诱导的通道活性及其对癌细胞行为的影响.

主要方法:

  • 单通道全细胞补丁电生理学研究通道活性.
  • 在K562细胞中进行 δ-ENaC 敲除 (KD),以评估子单元的参与.
  • 在体外的迁移和入侵测试以评估细胞运动性.
  • 使用血清蛋白酶抑制剂 (SBTI,α2-抗 плазмин) 来确认蛋白质分解活性的作用.

主要成果:

  • 在K562和U937白血病细胞系中,α-chymotrypsin和等离子素直接激活通道.
  • 降低δ-ENaC阻止了α-chymotrypsin诱导的通道激活,证实了δ-ENaC的作用.
  • α-chymotrypsin和等离子体显著降低K562细胞的迁移和入侵;这种效应被δ-ENaC KD部分取消.
  • α-chymotrypsin在U937和HL-60细胞中减少了迁移,但在缺乏功能ENAC的MOLT-4细胞中没有.
  • 蛋白酶抑制剂阻断了对细胞迁移/侵入的影响,表明需要蛋白质分解活性.

结论:

  • 细胞外血清蛋白酶α-chymotrypsin和等离子体激活白血病细胞中涉及δ-ENaC的通道.
  • 这些蛋白酶减少白血病细胞的迁移和入侵,这表明它们在癌症进展中起着调节作用.
  • 血清蛋白酶通过ENaC作为透性的调节剂,影响转化血细胞的运动性.