Role of phospholipases D1 and 2 in astroglial proliferation: effects of specific inhibitors and genetic deletion
Role of phospholipases D1 and 2 in astroglial proliferation: effects of specific inhibitors and genetic deletion
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DOI：
10.1016/j.ejphar.2015.05.004
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发表时间：
2015-08-15
影响因子：
5
通讯作者：
Klein, Jochen
中科院分区：
文献类型：
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作者：
Burkhardt, Ute;Beyer, Sandra;Klein, Jochen
磷脂酶D（OLD）活性与许多细胞类型（包括肿瘤细胞）的增殖有关。在本研究中，我们研究了PLD 1和PLD 2基因缺失和特定PLD 1和PLD 2抑制剂对原代小鼠星形胶质细胞中OLD活性和细胞增殖的影响。基础和刺激的OLD活性在PLD 1/2双敲除中可忽略不计。OLD活性显着降低PLD 1缺陷细胞时，胎牛血清（FCS），胰岛素样生长因子1（IGF-1）或佛波酯被用作兴奋剂。在佛波酯刺激的细胞中证实了500 nM的OLD抑制剂VU 0359595和VU 0285655 -1的特异性。在基础和刺激条件下，在OLD缺陷细胞系中观察到细胞增殖的显著减少。在500 nM时，PLD 1抑制剂VU 0359595降低了PLD 2缺陷细胞的增殖，但也降低了由IGF-1或佛波酯刺激的PLD 1缺陷细胞的增殖。反之亦然，在500 nM时，PLD 2抑制剂VU 0285655 -1降低了暴露于IGF-1的PLD 1-缺陷细胞中的增殖，但也降低了暴露于IGF-1的PLD 2-缺陷细胞中的增殖。在5 μ M的抑制剂显示非特异性的影响，因为它们抑制细胞增殖，即使在PLD 1/2双敲除。总之，OLD的抑制与PLD 1或PLD 2缺陷的星形胶质细胞中细胞增殖的减少平行发生。合成的OLD抑制剂在低（纳摩尔）浓度下显示出对OLD的高特异性，但在高（微摩尔）浓度下使用时对细胞增殖具有额外的非特异性作用。(C)2015 Elsevier B. V.版权所有。
Phospholipase D (OLD) activity has been linked to proliferation in many cell types including tumor cells. In the present study, we investigated the effects of genetic deletion of PLD1 and PLD2 and of specific PLD1 and PLD2 inhibitors on OLD activity and cell proliferation in primary mouse astrocytes. Basal and stimulated OLD activity was negligible in PLD1/2 double knockouts. OLD activity was significantly reduced in PLD1-deficient cells when fetal calf serum (FCS), insulin-like growth factor 1 (IGF-1) or phorbol ester was used as a stimulant. The specificity of OLD inhibitors VU0359595 and VU0285655-1 at 500 nM was confirmed in phorbol ester-stimulated cells. Significant reductions of cell proliferation were observed in OLD-deficient cell lines under basal and stimulated conditions. At 500 nM, the PLD1 inhibitor VU0359595 reduced proliferation in PLD2-deficient cells, but also in PLD1-deficient cells stimulated by IGF-1 or phorbol ester. Vice versa, at 500 nM, the PLD2 inhibitor VU0285655-1 reduced proliferation in PLD1-cleficient cells, but also in PLD2-deficient cells exposed to IGF-1. At 5 mu M both inhibitors showed non-specific effects because they inhibited cell proliferation even in PLD1/2 double knockouts. Summarizing, inhibition of OLD occurs in parallel with reduced cell proliferation in astrocytes which are deficient in PLD1 or PLD2. Synthetic OLD inhibitors show high specificity for OLD in low (nanomolar) concentrations, but have additional, non-specific effects on cell proliferation when used at high (micromolar) concentrations. (C) 2015 Elsevier B.V. All rights reserved.