Magnetic resonance spectroscopy monitoring of mitogen-activated protein kinase signaling inhibition
Magnetic resonance spectroscopy monitoring of mitogen-activated protein kinase signaling inhibition
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DOI：
10.1158/0008-5472.can-03-2981
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发表时间：
2005-04-15
期刊：
影响因子：
11.2
通讯作者：
Ronen, SM
中科院分区：
文献类型：
--
作者：
Beloueche-Babari, M;Jackson, LE;Ronen, SM
几种丝裂原活化蛋白激酶(MAPK)信号抑制剂目前正在进行临床试验，作为新的基于机制的抗癌治疗策略的一部分。本研究旨在利用磁共振波谱技术检测人乳腺癌细胞和结肠癌细胞中MAPK信号抑制的生物标志物。我们研究了原型MAPK激酶抑制剂U0126对MDA-MB-231、MCF-7和Hs578T乳腺癌细胞以及HCT116结肠癌细胞P-31-MR谱的影响。50mU·m o l/L U0126作用2、4、8、16、24、32、40h后，细胞外信号调节蛋白1/2的磷酸化水平从2小时起受到抑制。提取细胞的P-31-MR谱表明，这与磷胆碱水平在治疗后8小时显著下降至78+/-8%，16小时为74+/-8%，24小时为66+/-7%，32小时为71+/-10%，40小时为65-10%。而低浓度的10mU/L U0126作用40h，对细胞内P-ERK1/2和磷胆碱水平均无明显影响。用50mU·mol/L U0126去除MCF-7和Hs578T细胞中P-ERK1/2后，细胞内的磷胆碱水平分别下降到40小时的51+/-17%和48小时的23+/-12%。同样，在HCT116细胞中，30 mU·mol/L U0126可导致P-ERK1/2耗竭，磷胆碱水平在处理后16小时降至80+/-9%，24小时降至61+/-4%。在MDA-MB-231和HCT116细胞中，磷胆碱的减少与P-ERK1/2水平的下降呈正相关。我们的结果表明，用U0126抑制MAPK信号与细胞内磷胆碱水平的时间依赖性降低有关。因此，磷胆碱有可能成为监测MAPK信号阻断的非侵入性药效学标志物。
Several mitogen-activated protein kinase (MAPK) signaling inhibitors are currently undergoing clinical trial as part of novel mechanism-based anticancer treatment strategies. This study was aimed at detecting biomarkers of MAPK signaling inhibition in human breast and colon carcinoma cells using magnetic resonance spectroscopy. We investigated the effect of the prototype MAPK kinase inhibitor U0126 on the P-31-MR spectra of MDA-MB-231, MCF-7 and Hs578T breast, and HCT116 colon carcinoma cells. Treatment of MDA-MB-231 cells with 50 mu mol/L U0126 for 2, 4, 8, 16, 24, 32, and 40 hours caused inhibition of extracellular signal-regulated kinases (ERK1/2) phosphorylation from 2 hours onwards. P-31-MR spectra of extracted cells indicated that this was associated with a significant drop in phosphocholine levels to 78 +/- 8% at 8 hours, 74 +/- 8% at 16 hours, 66 +/- 7% at 24 hours, 71 +/- 10% at 32 hours, and 65 10% at 40 hours post-treatment. In contrast, the lower concentration of 10 mu mol/L U0126 for 40 hours had no significant effect on either P-ERK1/2 or phosphocholine levels in MDA-MB-231 cells. Depletion of P-ERK1/2 in MCF-7 and Hs578T cells with 50 mu mol/L U0126 also produced a drop in phosphocholine levels to 51 +/- 17% at 40 hours and 23 +/- 12% at 48 hours, respectively. Similarly, in HCT116 cells, inhibition with 30 mu mol/L U0126 caused depletion of P-ERK1/2 and a decrease in phosphocholine levels to 80 +/- 9% at 16 hours and 61 +/- 4% at 24 hours post-treatment. The reduction in phosphocholine in MDA-MB-231 and HCT116 cells correlated positively with the drop in P-ERK1/2 levels. Our results show that MAPK signaling inhibition with U0126 is associated with a time-dependent decrease in cellular phosphocholine levels. Thus, phosphocholine has potential as a noninvasive pharmacodynamic marker for monitoring MAPK signaling blockade.