Shih Yung-Luen, Chou Hsiao-Min, Chou Hsiu-Chen, Lu Hsu-Feng, Chu Yung-Lin, Shang Hung-Sheng, Chung Jing-Gung
Department of Pathology and Laboratory Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan.
Environ Toxicol. 2017 Sep;32(9):2097-2112. doi: 10.1002/tox.22417. Epub 2017 Apr 26.
Casticin, a polymethoxyflavone, is one of the major active components obtained from Fructus viticis, which have been shown to have anticancer activities including induce cell apoptosis in human cancer cells. The aim of this study was to investigate the molecular mechanisms by which casticin inhibits cell migration and invasion of mouse melanoma B16F10 cells. Cell viability was examined by MTT assay and the results indicated that casticin decreased the total percentages of viable cells in dose-dependent manners. Casticin affected cell migration and invasion in B16F10 cells were examined by wound healing mobility assay and Boyden chamber migration and invasion assay and results indicated that casticin inhibited cell migration and invasion in dose-dependent manners. Western blotting was used to examine the protein expression of B16F10 cells after exposed to casticin and the results showed that casticin decreased the expressions of MMP-9, MMP-2, MMP-1, FAK, 14-3-3, GRB2, Akt, NF-κB p65, SOS-1, p-EGFR, p-JNK 1/2, uPA, and Rho A in B16F10 cells. Furthermore, cDNA microarray assay was used to show that casticin affected associated gene expression of cell migration and invasion and the results indicated that casticin affected some of the gene expression such as increased SCN1B (cell adhesion molecule 1) and TIMP2 (TIMP metallopeptidase inhibitor 2) and decreased NDUFS4 (NADH dehydrogenase (ubiquinone) Fe-S protein4), VEGFA (vascular endothelial growth factor A), and DDIT3 (DNA-damage-inducible transcript 3) which associated cell migration and invasion in B16F10 cells. Based on those observations, we suggest that casticin could be used as a novel anticancer metastasis of melanoma cancer in the future.
紫花牡荆素是一种多甲氧基黄酮，是从蔓荆子中提取的主要活性成分之一，已被证明具有抗癌活性，包括诱导人类癌细胞凋亡。本研究的目的是探讨紫花牡荆素抑制小鼠黑色素瘤B16F10细胞迁移和侵袭的分子机制。采用MTT法检测细胞活力，结果表明紫花牡荆素以剂量依赖的方式降低了活细胞的总百分比。通过伤口愈合迁移试验、Boyden小室迁移和侵袭试验检测紫花牡荆素对B16F10细胞迁移和侵袭的影响，结果表明紫花牡荆素以剂量依赖的方式抑制细胞迁移和侵袭。采用蛋白质免疫印迹法检测紫花牡荆素作用后B16F10细胞的蛋白表达，结果显示紫花牡荆素降低了B16F10细胞中MMP-9、MMP-2、MMP-1、FAK、14-3-3、GRB2、Akt、NF-κB p65、SOS-1、p-EGFR、p-JNK 1/2、uPA和Rho A的表达。此外，利用cDNA微阵列分析表明紫花牡荆素影响细胞迁移和侵袭相关基因的表达，结果表明紫花牡荆素影响了一些基因的表达，如增加了SCN1B（细胞粘附分子1）和TIMP2（TIMP金属肽酶抑制剂2）的表达，降低了NDUFS4（NADH脱氢酶（泛醌）Fe-S蛋白4）、VEGFA（血管内皮生长因子A）和DDIT3（DNA损伤诱导转录物3）的表达，这些基因与B16F10细胞的迁移和侵袭有关。基于这些观察结果，我们认为紫花牡荆素未来可作为一种新型的黑色素瘤抗癌转移药物。
