Enterolactone alters FAK-Src signaling and suppresses migration and invasion of lung cancer cell lines.
Enterolactone alters FAK-Src signaling and suppresses migration and invasion of lung cancer cell lines.
复制标题
肠环酮改变FAK-SRC信号并抑制肺癌细胞系的迁移和侵袭。
DOI：
10.1186/s12906-016-1512-3
复制
发表时间：
2017-01-09
影响因子：
--
通讯作者：
Reindl KM
中科院分区：
文献类型：
--
作者：
Chikara S;Lindsey K;Borowicz P;Christofidou-Solomidou M;Reindl KM
化疗药物的全身毒性以及与靶向转移性肿瘤相关的挑战是当前肺癌治疗方法的限制因素。为了解决这些问题，人们对植物来源的生物活性成分的抗癌特性进行了研究，因为其中许多药物对健康组织无毒。肠内酯 (EL) 是一种源自亚麻籽的哺乳动物木酚素，已证明对多种癌症具有抗迁移特性，但 EL 尚未在肺癌中进行研究，其抗癌机制尚不明确。我们假设EL可以通过影响FAK-Src信号通路来抑制肺癌细胞的运动。通过进行 3-(4, 5-二甲基噻唑-2-基)-2, 5-脱苯基四唑溴化物 (MTT) 测定，确定了 EL 对 A549 和 H460 人肺癌细胞的无毒浓度。 EL 对肺癌细胞系的抗迁移和抗侵袭潜力是通过划痕愈合和 Matrigel® 侵袭测定来确定的。使用鬼笔环肽结合的罗丹明染料和荧光显微镜测定 EL 处理的细胞中丝状肌动蛋白 (F-肌动蛋白) 纤维密度和长度的变化。通过免疫细胞化学测定 EL 处理后粘着斑中的纽蛋白表达。使用 PCR 阵列、qRT-PCR 和蛋白质印迹法测定 EL 处理的肺癌细胞中 FAK-Src 信号分子的基因和蛋白质表达水平。无毒浓度的 EL 以浓度和时间依赖性方式抑制肺癌细胞迁移和侵袭。 EL治疗降低了肺癌细胞系中F-肌动蛋白纤维的密度和数量，并减少了粘着斑的数量和大小。 EL 降低了肺癌细胞中 FAK 及其下游靶标 Src、桩蛋白的磷酸化，并降低了细胞运动相关基因 RhoA、Rac1 和 Cdc42 的 mRNA 表达。我们的数据表明，EL 通过改变 FAK 活性以及随后激活粘着斑形成和细胞骨架重排所需的下游蛋白来抑制肺癌细胞的运动和侵袭。因此，给予EL可能作为抑制肺肿瘤细胞运动、侵袭和转移的安全且补充的方法。本文的在线版本 (doi:10.1186/s12906-016-1512-3) 包含补充材料，可供授权用户使用。
Systemic toxicity of chemotherapeutic agents and the challenges associated with targeting metastatic tumors are limiting factors for current lung cancer therapeutic approaches. To address these issues, plant-derived bioactive components have been investigated for their anti-cancer properties because many of these agents are non-toxic to healthy tissues. Enterolactone (EL) is a flaxseed-derived mammalian lignan that has demonstrated anti-migratory properties for various cancers, but EL has not been investigated in the context of lung cancer, and its anticancer mechanisms are ill-defined. We hypothesized that EL could inhibit lung cancer cell motility by affecting the FAK-Src signaling pathway. Non-toxic concentrations of EL were identified for A549 and H460 human lung cancer cells by conducting 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Dephenyltetrazolium Bromide (MTT) assays. The anti-migratory and anti-invasive potential of EL for lung cancer cell lines was determined by scratch wound healing and Matrigel® invasion assays. Changes in filamentous actin (F-actin) fiber density and length in EL-treated cells were determined using phalloidin-conjugated rhodamine dye and fluorescent microscopy. Vinculin expression in focal adhesions upon EL treatment was determined by immunocytochemistry. Gene and protein expression levels of FAK-Src signaling molecules in EL-treated lung cancer cells were determined using PCR arrays, qRT-PCR, and western blotting. Non-toxic concentrations of EL inhibited lung cancer cell migration and invasion in a concentration- and time-dependent manner. EL treatment reduced the density and number of F-actin fibers in lung cancer cell lines, and reduced the number and size of focal adhesions. EL decreased phosphorylation of FAK and its downstream targets, Src, paxillin, and decreased mRNA expression of cell motility-related genes, RhoA, Rac1, and Cdc42 in lung cancer cells. Our data suggest that EL suppresses lung cancer cell motility and invasion by altering FAK activity and subsequent activation of downstream proteins needed for focal adhesion formation and cytoskeletal rearrangement. Therefore, administration of EL may serve as a safe and complementary approach for inhibiting lung tumor cell motility, invasion, and metastasis. The online version of this article (doi:10.1186/s12906-016-1512-3) contains supplementary material, which is available to authorized users.