Inhibition of p38 MAPK activity leads to cell type-specific effects on the molecular circadian clock and time-dependent reduction of glioma cell invasiveness.
Inhibition of p38 MAPK activity leads to cell type-specific effects on the molecular circadian clock and time-dependent reduction of glioma cell invasiveness.
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DOI：
10.1186/s12885-017-3896-y
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发表时间：
2018-01-10
期刊：
影响因子：
3.8
通讯作者：
Bell-Pedersen D
中科院分区：
文献类型：
--
作者：
Goldsmith CS;Kim SM;Karunarathna N;Neuendorff N;Toussaint LG;Earnest DJ;Bell-Pedersen D
生物钟是真核生物保持生物时间的基础。生物钟机制依赖于生物化学信号通路来检测环境刺激，并调节整个身体中时钟控制基因的表达。MAPK信号通路在哺乳动物的昼夜节律输入和输出通路中起作用，这取决于组织;然而，对p38 MAPK（一种已建立的肿瘤抑制因子）在哺乳动物昼夜节律系统中的作用知之甚少。p38 MAPK的表达和活性增加与癌症（包括多形性胶质母细胞瘤）的不良预后相关;然而，p38 MAPK抑制剂的毒性限制了其临床应用。在这里，我们测试，如果定时应用特定的p38 MAPK抑制剂VX-745减少胶质瘤细胞的体外侵袭性。蛋白质印迹法检测不同细胞系中活性磷酸化p38 MAPK的水平和节律性积累。来自时钟基因荧光素酶报告细胞系的节律荧光素酶活性用于测试p38 MAPK抑制对时钟性质的影响，如使用阻尼正弦拟合和Levenberg-Marquardt算法所确定的。非线性回归和Akaike的信息标准被用来建立节律性。Boyden室测定用于测量用VX-745进行一天中特定时间处理后的胶质瘤细胞侵袭性。使用t检验确定显著差异。我们证明了在小鼠成纤维细胞和SCN细胞系中，p38 MAPK周期在时钟控制下的活性。磷酸化p38 MAPK的水平在生物钟缺陷细胞中显著降低，表明生物钟在该途径的激活中起重要作用。用VX-745抑制p38 MAPK活性导致分子钟中细胞类型特异性周期变化。此外，磷酸化p38 MAPK水平在HA胶质细胞中是节律性的，而在侵袭性IM 3胶质瘤细胞中是高水平的且是低水平的。我们表明，在一天中的时间，当在HA细胞中的水平通常较低时，在生物钟的控制下，抑制IM 3细胞中的p38 MAPK活性，显着降低了IM 3的侵袭性。如果在一天中的适当时间给药，用p38 MAPK抑制剂治疗胶质瘤可能更有效，毒性更小。本文的在线版本（10.1186/s12885-017-3896-y）包含补充材料，可供授权用户使用。
The circadian clock is the basis for biological time keeping in eukaryotic organisms. The clock mechanism relies on biochemical signaling pathways to detect environmental stimuli and to regulate the expression of clock-controlled genes throughout the body. MAPK signaling pathways function in both circadian input and output pathways in mammals depending on the tissue; however, little is known about the role of p38 MAPK, an established tumor suppressor, in the mammalian circadian system. Increased expression and activity of p38 MAPK is correlated with poor prognosis in cancer, including glioblastoma multiforme; however, the toxicity of p38 MAPK inhibitors limits their clinical use. Here, we test if timed application of the specific p38 MAPK inhibitor VX-745 reduces glioma cell invasive properties in vitro. The levels and rhythmic accumulation of active phosphorylated p38 MAPK in different cell lines were determined by western blots. Rhythmic luciferase activity from clock gene luciferase reporter cells lines was used to test the effect of p38 MAPK inhibition on clock properties as determined using the damped sine fit and Levenberg–Marquardt algorithm. Nonlinear regression and Akaike’s information criteria were used to establish rhythmicity. Boyden chamber assays were used to measure glioma cell invasiveness following time-of-day-specific treatment with VX-745. Significant differences were established using t-tests. We demonstrate the activity of p38 MAPK cycles under control of the clock in mouse fibroblast and SCN cell lines. The levels of phosphorylated p38 MAPK were significantly reduced in clock-deficient cells, indicating that the circadian clock plays an important role in activation of this pathway. Inhibition of p38 MAPK activity with VX-745 led to cell-type-specific period changes in the molecular clock. In addition, phosphorylated p38 MAPK levels were rhythmic in HA glial cells, and high and arrhythmic in invasive IM3 glioma cells. We show that inhibition of p38 MAPK activity in IM3 cells at the time of day when the levels are normally low in HA cells under control of the circadian clock, significantly reduced IM3 invasiveness. Glioma treatment with p38 MAPK inhibitors may be more effective and less toxic if administered at the appropriate time of the day. The online version of this article (10.1186/s12885-017-3896-y) contains supplementary material, which is available to authorized users.
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影响因子：
3.5
作者：
Ko ML;Shi L;Tsai JY;Young ME;Neuendorff N;Earnest DJ;Ko GY
通讯作者：
Ko GY
影响因子：
11.4
作者：
Bulavin, DV;Saito, S;Fornace, AJ
通讯作者：
Fornace, AJ
影响因子：
9.2
作者：
Cermakian, N;Pando, MP;Sassone-Corsi, P
通讯作者：
Sassone-Corsi, P
DOI：
10.1073/pnas.88.9.3720
发表时间：
1991-05-01
影响因子：
11.1
作者：
HAI, T;CURRAN, T
通讯作者：
CURRAN, T
影响因子：
5.7
作者：
Demuth, Tim;Reavie, Linsey B.;Berens, Michael E.
通讯作者：
Berens, Michael E.