Ling Tao, Wang Wei, Hu Chen, Yan Xin, Xu Yu-Ting, Tang Chun, Kou Yu, Liu Liang
Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou 225001, China.
Zhongguo Zhong Yao Za Zhi. 2018 Dec;43(23):4718-4723. doi: 10.19540/j.cnki.cjcmm.20180710.001.
The purpose of this study was to investigate the inhibitory effect of the main 9,10-dihydrophenanthrene orchinol isolated from Spiranthes sinensis Radix et Herba on the invasion and migration of human gastric cancer SGC-7901 cells and its preliminary molecular mechanism. SGC-7901 cells were cultured in vitro, after the cells were treated with different final concentrations(5, 10, 20, 40, 80 μmol·L⁻¹) of orchinol for 24, 48 or 72 hours, the effect of orchinol on cell viability was measured by MTT assay. Wound healing and Transwell assays were performed to determine the effects of different final concentrations(5, 10, 20, 40 μmol·L⁻¹) of orchinol for 48 hour on invasion and migration abilities of SGC-7901 cells, respectively. The protein expression levels of β-catenin, Wnt-3α, DvL2, cyclinD1 and GSK-3β were detected by Western blot. The results showed that 5-80 μmol·L⁻¹ orchinol inhibited the viability of SGC-7901 cells in a dose-dependent and time-dependent manner, and the IC₅₈ values of 24, 48 and 72 hours were 77.79, 42.96 and 7.85 μmol·L⁻¹, respectively. Compared with the control group, the ability of invasion and migration of SGC-7901 cells was significantly inhibited after treated with 5, 10 and 20 μmol·L⁻¹ orchinol for 48 hours (<0.05, <0.01), and the dose-effect relationship was observed. The results of Western blot showed that orchinol could significantly down-regulate the protein expression levels of β-catenin, Wnt3a, DvL2 and cyclinD1, and up-regulate the protein expression level of GSK-3β(<0.05, <0.01, <0.001). The above results suggest that orchinol can obviously inhibit the invasion and migration of SGC-7901 cells, which may be related to its inhibition of Wnt3a/β-catenin signaling pathway and the proteins expression of downstream genes.
本研究旨在探讨从绶草中分离得到的主要成分9,10-二氢菲山奈酚对人胃癌SGC-7901细胞侵袭和迁移的抑制作用及其初步分子机制。体外培养SGC-7901细胞，用不同终浓度（5、10、20、40、80 μmol·L⁻¹）的山奈酚处理细胞24、48或72小时后，采用MTT法检测山奈酚对细胞活力的影响。分别进行划痕愈合实验和Transwell实验，以确定不同终浓度（5、10、20、40 μmol·L⁻¹）的山奈酚作用48小时对SGC-7901细胞侵袭和迁移能力的影响。通过蛋白质免疫印迹法检测β-连环蛋白、Wnt-3α、DvL2、细胞周期蛋白D1和糖原合成酶激酶-3β的蛋白表达水平。结果显示，5 - 80 μmol·L⁻¹山奈酚以剂量和时间依赖性方式抑制SGC-7901细胞活力，24、48和72小时的IC₅₀值分别为77.79、42.96和7.85 μmol·L⁻¹。与对照组相比，5、10和20 μmol·L⁻¹山奈酚处理48小时后，SGC-7901细胞的侵袭和迁移能力显著受到抑制（<0.05，<0.01），呈现剂量-效应关系。蛋白质免疫印迹结果表明，山奈酚可显著下调β-连环蛋白、Wnt3a、DvL2和细胞周期蛋白D1的蛋白表达水平，并上调糖原合成酶激酶-3β的蛋白表达水平（<0.05，<0.01，<0.001）。上述结果表明，山奈酚可明显抑制SGC-7901细胞的侵袭和迁移，这可能与其抑制Wnt3a/β-连环蛋白信号通路及下游基因的蛋白表达有关。
