Differential protein expression and novel biomarkers related to 5-FU resistance in a 3D colorectal adenocarcinoma model
Differential protein expression and novel biomarkers related to 5-FU resistance in a 3D colorectal adenocarcinoma model
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DOI：
10.3892/or.2014.3337
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发表时间：
2014-10-01
期刊：
影响因子：
4.2
通讯作者：
Kuh, Hyo-Jeong
中科院分区：
文献类型：
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作者：
Lee, Sang-Hak;Nam, Jae Kook;Kuh, Hyo-Jeong
多细胞类球体(MCS)是一种反映体内微环境条件，高度代表实体瘤无血管区域的体外模型。因此，它被认为是研究耐药的最合适的模型。我们使用DLD-1细胞比较了3D MCS模型和2D单层细胞对5-FU(5-FU)的化疗敏感性和差异蛋白表达。通过蛋白质印迹分析，我们分析了几个重要的信号分子。尽管根据样本的不同，我们观察到了表达水平的不同变化，但我们没有得到显著的发现。值得注意的是，5-FU暴露后，单层细胞的p-mTOR水平降低，而MCSs的p-mTOR水平较高。通过2-DE和MALDI-TOF分析鉴定了9个新的蛋白质，它们在MCS模型和单层之间表现出差异表达。其中，崩解素反应介质蛋白2(CRMP-2)、DNA复制复合体gins蛋白PSF2(PSF-2)和硒结合蛋白1(SBP-1)不仅表达差异显著，而且在5-FU暴露后表达下降，提示它们可能作为5-FU敏感性(CRMP-2、PSF-2)和耐药(SBP-1)的新生物标志物。总体而言，本研究表明，与单层相比，MCSs培养的人结直肠癌细胞对5-FU的耐药性更强，并确定p-mTOR、CRMP-2、PSF-2和SBP-1是结直肠癌5-FU化疗敏感性/耐药性的新的潜在生物标志物，这些发现值得进一步研究。
The multicellular spheroid (MCS) is an in vitro model which is highly representative of the avascular region of solid tumors by reflecting microenvironmental conditions in vivo. Hence, it is considered the most appropriate model for studying drug resistance. We compared chemosensitivity to 5-fluorouracil (5-FU) and differential protein expression between the 3D MCS model and the 2D monolayers using DLD-1 cells. We analyzed several significant signaling molecules through western blot analysis. Although various changes in the expression level were observed depending on the samples, we did not obtained remarkable findings. Notably, the level of p-mTOR decreased upon 5-FU exposure in the monolayers, while its level was higher in the MCSs. Nine novel proteins were identified by 2-DE and MALDI-TOF analysis for exhibiting differential expression between the MCS model and the monolayers. Among these, collapsin response mediator protein 2 (CRMP-2), DNA replication complex GINS protein PSF2 (PSF-2) and selenium-binding protein 1 (SBP-1) were notable not only for their differential expression but also for decreased expression following 5-FU exposure, indicating their possible roles as novel biomarkers for sensitivity (CRMP-2, PSF-2) as well as resistance (SBP-1) to 5-FU. Overall, the present study demonstrated greater 5-FU resistance in human colorectal cancer cells grown as MCSs compared to monolayers and identified p-mTOR, CRMP-2, PSF-2 and SBP-1 as novel potential biomarkers of 5-FU chemosensitivity/resistance for human colorectal cancer, findings which warrant further investigation.