Clinical Institute of Pathology, Medical University of Vienna, Austria.
Department of Clinical Pharmacy and Diagnostics, University of Vienna, A-1090 Vienna, Austria.
Food Chem Toxicol. 2018 Jan;111:114-124. doi: 10.1016/j.fct.2017.11.015. Epub 2017 Nov 10.
Mechanisms how colorectal cancer (CRC) cells penetrate blood micro-vessel endothelia and metastasise is poorly understood. To study blood endothelial cell (BEC) barrier breaching by CRC emboli, an in vitro assay measuring BEC-free areas underneath SW620 cell spheroids, so called "circular chemorepellent induced defects" (CCIDs, appearing in consequence of endothelial retraction), was adapted and supported by Western blotting, EIA-, EROD- and luciferase reporter assays. Inhibition of ALOX12 or NF-κB in SW620 cells or BECs, respectively, caused attenuation of CCIDs. The FDA approved drugs vinpocetine [inhibiting ALOX12-dependent 12(S)-HETE synthesis], ketotifen [inhibiting NF-κB], carbamazepine and fenofibrate [inhibiting 12(S)-HETE and NF-κB] significantly attenuated CCID formation at low μM concentrations. In the 5-FU-resistant SW620-R/BEC model guanfacine, nifedipine and proadifen inhibited CCIDs stronger than in the naïve SW620/BEC model. This indicated that in SW620-R cells formerly silent (yet unidentified) genes became expressed and targetable by these drugs in course of resistance acquisition. Fenofibrate, and the flavonoids hispidulin and apigenin, which are present in medicinal plants, spices, herbs and fruits, attenuated CCID formation in both, naïve- and resistant models. As FDA-approved drugs and food-flavonoids inhibited established and acquired intravasative pathways and attenuated BEC barrier-breaching in vitro, this warrants testing of these compounds in CRC models in vivo.
结直肠癌（CRC）细胞穿透血液微血管内皮并转移的机制尚不清楚。为了研究 CRC 栓子对血液内皮细胞（BEC）屏障的破坏，我们对一种体外测定法进行了适应性改进，该方法可测量 SW620 细胞球体下方无 BEC 的区域，即所谓的“圆形趋化抑制诱导缺陷”（CCIDs，由于内皮细胞收缩而出现），并通过 Western blot、EIA、EROD 和荧光素酶报告基因测定法进行了支持。在 SW620 细胞或 BEC 中分别抑制 ALOX12 或 NF-κB，可导致 CCIDs 减弱。美国食品和药物管理局批准的药物长春西汀（抑制 ALOX12 依赖性 12(S)-HETE 合成）、酮替芬（抑制 NF-κB）、卡马西平和非诺贝特（抑制 12(S)-HETE 和 NF-κB）在低 μM 浓度下显著减弱 CCID 的形成。在 5-FU 耐药的 SW620-R/BEC 模型中，胍法辛、硝苯地平和普罗地芬比在未耐药的 SW620/BEC 模型中抑制 CCIDs 的作用更强。这表明，在 SW620-R 细胞中，以前沉默（但未鉴定）的基因在获得耐药性的过程中可被这些药物表达并作为靶点。非诺贝特以及存在于药用植物、香料、草药和水果中的类黄酮荭草苷和芹菜素，在两种模型中均减弱了 CCID 的形成。由于美国食品和药物管理局批准的药物和食物类黄酮抑制了已建立的和获得的血管内途径，并减弱了体外 BEC 屏障的破坏，因此有必要在 CRC 动物模型中测试这些化合物。
