With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volume
With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volume
复制标题
DOI：
10.1152/ajpcell.00203.2011
复制
发表时间：
2011-11-01
影响因子：
5.5
通讯作者：
Sontheimer, Harald
中科院分区：
文献类型：
--
作者：
Haas, Brian R.;Cuddapah, Vishnu A.;Sontheimer, Harald
哈斯BR，Cuddapah VA，Watkins S，Rohn KJ，Dy TE，Sontheimer H.无赖氨酸激酶3（WNK 3）通过调节细胞体积刺激胶质瘤侵袭。美国生理学杂志细胞生理学301：C1150-C1160，2011年。首次发表于2011年8月3日; doi：10.1152/ajpcell.00203.2011。在最常见和最致命的原发性脑肿瘤中，高级别胶质瘤通过弥漫性侵入周围脑实质而逃避完全手术切除。通过紧密的细胞外空间导航需要入侵胶质瘤细胞来改变它们的形状和体积。细胞体积的变化是通过渗透物沿着与义务水的跨膜转运实现的。钠-钾-氯化物协同转运体亚型-1（NKCC 1）在这一过程中起着关键作用，先前的工作已经证明，NKCC 1抑制通过干扰所需的细胞体积变化而在体外和体内损害胶质瘤侵袭。在这项研究中，我们发现神经胶质瘤中的NKCC 1活性需要无赖氨酸激酶-3（WNK 3）激酶。患者活检和患者来源的细胞系的蛋白质印迹显示Ste-20相关的富含脯氨酸的激酶（SPAK）、氧化应激反应激酶（OSR 1）和WNK家族成员1、3和4的显著表达。其中，只有WNK 3与NKCC 1在细胞体积变化时共定位和共免疫沉淀。使用特异性短发夹RNA构建体稳定敲除WNK 3完全消除了NKCC 1活性，如通过布美他尼敏感性细胞体积调节的丧失所测量的。因此，WNK 3敲除细胞显示出降低的侵入跨Transwell屏障的能力，并且缺乏布美他尼敏感性迁移。这些数据表明WNK 3是NKCC 1的重要调节因子，并且WNK 3激活NKCC 1介导的离子转运，这是与细胞侵袭相关的细胞体积变化所必需的。
Haas BR, Cuddapah VA, Watkins S, Rohn KJ, Dy TE, Sontheimer H. With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volume. Am J Physiol Cell Physiol 301: C1150-C1160, 2011. First published August 3, 2011; doi:10.1152/ajpcell.00203.2011.-Among the most prevalent and deadly primary brain tumors, high-grade gliomas evade complete surgical resection by diffuse invasion into surrounding brain parenchyma. Navigating through tight extracellular spaces requires invading glioma cells to alter their shape and volume. Cell volume changes are achieved through transmembrane transport of osmolytes along with obligated water. The sodium-potassium-chloride cotransporter isoform-1 (NKCC1) plays a pivotal role in this process, and previous work has demonstrated that NKCC1 inhibition compromises glioma invasion in vitro and in vivo by interfering with the required cell volume changes. In this study, we show that NKCC1 activity in gliomas requires the With-No-Lysine Kinase-3 (WNK3) kinase. Western blots of patient biopsies and patient-derived cell lines shows prominent expression of Ste-20-related, prolinealanine-rich kinase (SPAK), oxidative stress response kinase (OSR1), and WNK family members 1, 3, and 4. Of these, only WNK3 colocalized and coimmunoprecipitated with NKCC1 upon changes in cell volume. Stable knockdown of WNK3 using specific short hairpin RNA constructs completely abolished NKCC1 activity, as measured by the loss of bumetanide-sensitive cell volume regulation. Consequently, WNK3 knockdown cells showed a reduced ability to invade across Transwell barriers and lacked bumetanide-sensitive migration. This data indicates that WNK3 is an essential regulator of NKCC1 and that WNK3 activates NKCC1-mediated ion transport necessary for cell volume changes associated with cell invasion.