The human ion channel TRPM2 modulates neuroblastoma cell survival and mitochondrial function through Pyk2, CREB, and MCU activation.
The human ion channel TRPM2 modulates neuroblastoma cell survival and mitochondrial function through Pyk2, CREB, and MCU activation.
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DOI：
10.1152/ajpcell.00098.2018
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发表时间：
2018-10-01
期刊：
影响因子：
--
通讯作者：
Miller BA
中科院分区：
文献类型：
--
作者：
Hirschler-Laszkiewicz I;Chen SJ;Bao L;Wang J;Zhang XQ;Shanmughapriya S;Keefer K;Madesh M;Cheung JY;Miller BA
瞬时受体电位melastatin通道亚家族成员2（TRPM 2）在细胞存活中具有重要功能，并且在许多癌症中高度表达。通过用CRISPR技术耗竭或表达显性阴性TRPM 2-S来抑制神经母细胞瘤中的TRPM 2已显示显著降低细胞活力。在这里，富含脯氨酸的酪氨酸激酶2（Pyk 2）在TRPM 2调节神经母细胞瘤活力的作用进行了探讨。在TRPM 2耗尽的细胞中，Pyk 2和cAMP反应元件结合蛋白（CREB）（下游靶点）的磷酸化和表达在应用化疗剂多柔比星后显著降低。野生型Pyk 2的过表达拯救了细胞活力。用shRNA减少Pyk 2表达降低细胞活力和CREB磷酸化和表达，证明Pyk 2调节CREB活化。TRPM 2缺失损害Pyk 2激活剂Src的磷酸化，这可能是减少Pyk 2磷酸化的机制。TRPM 2抑制先前被证明降低线粒体功能。在这里，CREB，Pyk 2和磷酸化的Src在TRPM 2耗尽细胞的线粒体中减少，与它们在调节线粒体蛋白的表达和活化中的作用一致。磷酸化Src和磷酸化和总CREB减少TRPM 2耗尽的细胞核。磷酸化Pyk 2和CREB的靶点线粒体钙单向转运体（MCU）的表达和功能显著降低。野生型TRPM 2而不是Ca 2+不渗透突变体E960 D在暴露于阿霉素的TRPM 2耗尽细胞中重建Pyk 2和CREB的磷酸化和表达。结果表明，TRPM 2表达通过Src，Pyk 2，CREB和MCU激活保护神经母细胞瘤的活力，这些激活在维持线粒体功能和细胞生物能量学中起关键作用。
Transient receptor potential melastatin channel subfamily member 2 (TRPM2) has an essential function in cell survival and is highly expressed in many cancers. Inhibition of TRPM2 in neuroblastoma by depletion with CRISPR technology or expression of dominant negative TRPM2-S has been shown to significantly reduce cell viability. Here, the role of proline-rich tyrosine kinase 2 (Pyk2) in TRPM2 modulation of neuroblastoma viability was explored. In TRPM2-depleted cells, phosphorylation and expression of Pyk2 and cAMP-responsive element-binding protein (CREB), a downstream target, were significantly reduced after application of the chemotherapeutic agent doxorubicin. Overexpression of wild-type Pyk2 rescued cell viability. Reduction of Pyk2 expression with shRNA decreased cell viability and CREB phosphorylation and expression, demonstrating Pyk2 modulates CREB activation. TRPM2 depletion impaired phosphorylation of Src, an activator of Pyk2, and this may be a mechanism to reduce Pyk2 phosphorylation. TRPM2 inhibition was previously demonstrated to decrease mitochondrial function. Here, CREB, Pyk2, and phosphorylated Src were reduced in mitochondria of TRPM2-depleted cells, consistent with their role in modulating expression and activation of mitochondrial proteins. Phosphorylated Src and phosphorylated and total CREB were reduced in TRPM2-depleted nuclei. Expression and function of mitochondrial calcium uniporter (MCU), a target of phosphorylated Pyk2 and CREB, were significantly reduced. Wild-type TRPM2 but not Ca2+-impermeable mutant E960D reconstituted phosphorylation and expression of Pyk2 and CREB in TRPM2-depleted cells exposed to doxorubicin. Results demonstrate that TRPM2 expression protects the viability of neuroblastoma through Src, Pyk2, CREB, and MCU activation, which play key roles in maintaining mitochondrial function and cellular bioenergetics.
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影响因子：
64.5
作者：
Cárdenas C;Miller RA;Smith I;Bui T;Molgó J;Müller M;Vais H;Cheung KH;Yang J;Parker I;Thompson CB;Birnbaum MJ;Hallows KR;Foskett JK
通讯作者：
Foskett JK
影响因子：
4.8
作者：
Chen, Shu-jen;Hoffman, Nicholas E.;Miller, Barbara A.
通讯作者：
Miller, Barbara A.
影响因子：
3.4
作者：
Bai, Ji-Zhong;Lipski, Janusz
通讯作者：
Lipski, Janusz
影响因子：
5.3
作者：
Giannoni, E;Buricchi, F;Chiarugi, P
通讯作者：
Chiarugi, P
影响因子：
16
作者：
Hara, Y;Wakamori, M;Mori, Y
通讯作者：
Mori, Y