Deguelin Impairs Cell Adhesion, Migration and Invasion of Human Lung Cancer Cells through the NF-κB Signaling Pathways
Deguelin Impairs Cell Adhesion, Migration and Invasion of Human Lung Cancer Cells through the NF-κB Signaling Pathways
复制标题
DOI：
10.1142/s0192415x1850012x
复制
发表时间：
2018-01-01
影响因子：
5.7
通讯作者：
Chung, Jing-Gung
中科院分区：
文献类型：
--
作者：
Hsiao, Yung-Ting;Fan, Ming-Jen;Chung, Jing-Gung
鱼藤素是从天然植物中分离得到的一种鱼藤素，具有抗肿瘤等生物活性。在本研究中，我们观察了鱼藤素对NCI-H292人肺癌细胞体外黏附、迁移和侵袭的影响。用流式细胞仪分析细胞存活率。采用细胞-基质粘附法测定细胞粘附率。创面愈合实验检测细胞迁移情况。用Boyden小室实验研究细胞的迁移和侵袭。用Western blotting和激光共聚焦显微镜检测蛋白质的表达。用凝胶电泳法检测核因子-kappaBp65与DNA的结合。我们选择0、0.5、1.0、1.5、2.0和2.5mU M的鱼藤素，发现这些浓度的鱼藤素对NCIH292细胞没有明显的细胞毒作用。因此，我们选择了这些浓度的鱼腥草素进行转移实验。我们发现鱼腥草素以剂量依赖的方式抑制细胞的黏附、迁移和侵袭，分别用伤口愈合和透气法检测。Dguelin下调NCI-H292细胞中MMP2/-9、SOS 1、Rho A、p-AKT(308)、p-ERK1/2、p-p38、p-JNK、NF-kappaB(65)和uPA的表达。激光共聚焦显微镜显示，鱼藤素抑制NCI-H292细胞胞核中PI3K、SOS-1、NF-kappa B(P65)的表达，但对PKC和RAS无明显影响。提示鱼藤素可作为一种新的抗肺癌转移药物。
Deguelin, a rotenoid, is isolated from a natural plant species, and has biological activities including antitumor function. In the present study, we investigated the effect of deguelin on the cell adhesion, migration and invasion of NCI-H292 human lung cancer cells in vitro. Cell viability was analyzed by using flow cytometer. Cell adhesion was determined by using the cell-matrix adhesion assay. Wound healing assay was used to examine cell migration. Cell migration and invasion were investigated using a Boyden chamber assay. The protein expression was measured by Western blotting and confocal laser microscopy. The electrophoretic mobility shift assay was used to measure NF-kappa B p65 binding to DNA. We selected the concentrations of deguelin at 0, 0.5, 1.0, 1.5, 2.0 and 2.5 mu M and we found that those concentrations of deguelin did not induce significant cytotoxic effects on NCIH292 cells. Thus, we selected those concentrations of deguelin for metastasis assay. We found that deguelin inhibited cell adhesion, migration and invasion in dose-dependent manners that was assayed by wound healing and transwell methods, respectively. Deguelin decreased the expression of MMP-2/-9, SOS 1, Rho A, p-AKT (308), p-ERK1/2, p-p38, p- JNK, NF-kappa B (65) and uPA in NCI- H292 cells. Deguelin suppressed the expression of PI3K, SOS 1, NF-kappa B(p65), but did not significantly affect PKC and Ras in the nuclei of NCI-H292 cells that were confirmed by confocal laser microscopy. We suggest that deguelin may be used as a novel anticancer metastasis of lung cancer in the future.