The Human Transient Receptor Potential Melastatin 2 Ion Channel Modulates ROS Through Nrf2
The Human Transient Receptor Potential Melastatin 2 Ion Channel Modulates ROS Through Nrf2
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DOI：
10.1038/s41598-019-50661-8
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发表时间：
2019-10-01
影响因子：
4.6
通讯作者：
Miller, Barbara A.
中科院分区：
文献类型：
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作者：
Bao, Lei;Festa, Fernanda;Miller, Barbara A.
瞬时受体电位melastatin通道亚家族成员2（TRPM 2）通过调节氧化应激在保护细胞活力中具有重要作用。TRPM 2在癌症中高度表达。当TRPM 2被抑制时，线粒体功能障碍，ROS水平增加，细胞活力降低。在此，探讨了NF-E2相关因子（Nrf 2）在TRPM 2介导的氧化应激抑制中的重要性。在TRPM 2耗竭的细胞中，抗氧化辅因子谷胱甘肽、NADPH和NADH显著减少。Nrf 2和IQGAP 1（一种受细胞内钙调节的Nrf 2稳定性调节剂）的细胞质和细胞核表达降低。受Nrf 2转录调节并参与GSH、NADPH和NADH生成的抗氧化酶显著降低，包括PRX 1和PRX 3、GPX 4、GSTP 1、GCLC和MTHFD 2。导致GSH产生的谷氨酰胺途径受到抑制，ATP和GTP水平受损。用野生型TRPM 2或Nrf 2而不是TRPM 2孔突变体E960 D重建，挽救了Nrf 2下游酶的表达并恢复了GSH和GTP。细胞活力、ROS、NADPH、NADH和ATP水平被TRPM 2完全拯救，部分被Nrf 2拯救。这些数据表明，TRPM 2通过调节Nrf 2调节的抗氧化途径和辅因子来维持氧化应激后的细胞存活。
Transient receptor potential melastatin channel subfamily member 2 (TRPM2) has an essential role in protecting cell viability through modulation of oxidative stress. TRPM2 is highly expressed in cancer. When TRPM2 is inhibited, mitochondria are dysfunctional, ROS levels are increased, and cell viability is reduced. Here, the importance of NF-E2-related factor (Nrf2) in TRPM2-mediated suppression of oxidant stress was explored. In TRPM2 depleted cells, antioxidant cofactors glutathione, NADPH, and NADH were significantly reduced. Cytoplasmic and nuclear expression of Nrf2 and of IQGAP1, a modulator of Nrf2 stability regulated by intracellular calcium, were decreased. Antioxidant enzymes transcriptionally regulated by Nrf2 and involved in GSH, NADPH, and NADH generation were significantly lower including PRX1 and PRX3, GPX4, GSTP1, GCLC, and MTHFD2. The glutamine pathway leading to GSH production was suppressed, and ATP and GTP levels were impaired. Reconstitution with wild type TRPM2 or Nrf2, but not TRPM2 pore mutant E960D, rescued expression of enzymes downstream of Nrf2 and restored GSH and GTP. Cell viability, ROS, NADPH, NADH, and ATP levels were fully rescued by TRPM2 and partially by Nrf2. These data show that TRPM2 maintains cell survival following oxidative stress through modulation of antioxidant pathways and cofactors regulated by Nrf2.