State Key Laboratory of Biocontrol, Sun Yat-sen (Zhongshan) University, Guangzhou, Guangdong, PR China.
Basic Medical College Jiangxi University of traditional Chinese Medicine, Nanchang, Jiangxi, PR China.
Phytomedicine. 2016 May 15;23(5):491-7. doi: 10.1016/j.phymed.2016.02.019. Epub 2016 Mar 3.
Ganoderma lucidum (G. lucidum) is an oriental medical mushroom that has been widely used in Asian countries for centuries to prevent and treat different diseases, including cancer.
HYPOTHESIS/PURPOSE: The objective of this study was to investigate the effect of A supercritical-CO2 extract of G. lucidum spores on the transforming growth factor beta 1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of cholangiocarcinoma cells.
This was an in vitro study with human cholangiocarcinoma TFK-1 cells treated with varying concentrations of G. lucidum.
A supercritical-CO2 extract of G. lucidum spores (GLE) was obtained from completely sporoderm-broken germinating G. lucidum spores by supercritical fluid carbon dioxide (SCF-CO2) extraction. GLE pre-incubated with human cholangiocarcinoma TFK-1 cells prior to TGF-β1 treatment (2ng/ml) for 48h. Changes in EMT markers were analyzed by western blotting and immunofluorescence. The formation of F-actin stress fibers was assessed via immunostaining with phalloidin and examined using confocal microscopy. Additionally, the effect of the GLE on TGF-β1-induced migration was investigated by a Boyden chamber assay.
TGF-β1-induced reduction in E-cadherin expression was associated with a loss of epithelial morphology and cell-cell contact. Concomitant increases in N-cadherin and Fibronectin were evident in predominantly elongated fibroblast-like cells. The GLE suppressed the TGF-β1-induced morphological changes and the changes in cadherin expression, and also inhibited the formation of F-actin stress fibers, which are a hallmark of EMT. The GLE also inhibited TGF-β1-induced migration of TFK-1 cells.
Our findings provide new evidence that GLE suppress cholangiocarcinoma migration in vitro through inhibition of TGF-β1-induced EMT. The GLE may be clinically applied in the prevention and/or treatment of cancer metastasis.
灵芝（Ganoderma lucidum）是一种药用蘑菇，在亚洲国家已有数百年的历史，被广泛用于预防和治疗各种疾病，包括癌症。
假设/目的：本研究旨在探讨灵芝孢子超临界-CO2 提取物（GLE）对转化生长因子-β1（TGF-β1）诱导的胆管癌细胞上皮-间充质转化（EMT）的影响。
这是一项体外研究，使用不同浓度 GLE 处理人胆管癌细胞 TFK-1。
采用超临界流体 CO2（SCF-CO2）萃取法从完全破壁萌发的灵芝孢子中提取灵芝孢子超临界-CO2 提取物（GLE）。GLE 与人类胆管癌细胞 TFK-1 孵育后，再用 TGF-β1（2ng/ml）处理 48h。采用 Western blot 和免疫荧光法分析 EMT 标志物的变化。用鬼笔环肽免疫染色检测 F-肌动蛋白应力纤维的形成，并通过共聚焦显微镜观察。此外，通过 Boyden 室测定研究 GLE 对 TGF-β1 诱导的迁移的影响。
TGF-β1 诱导的 E-钙黏蛋白表达减少与上皮形态和细胞间接触丧失有关。N-钙黏蛋白和纤连蛋白的表达增加，主要表现为拉长的成纤维细胞样细胞。GLE 抑制了 TGF-β1 诱导的形态变化和钙黏蛋白表达的变化，并抑制了 EMT 的标志性特征 F-肌动蛋白应力纤维的形成。GLE 还抑制了 TGF-β1 诱导的 TFK-1 细胞迁移。
我们的研究结果提供了新的证据，表明 GLE 通过抑制 TGF-β1 诱导的 EMT 来抑制胆管癌细胞的体外迁移。GLE 可能在癌症转移的预防和/或治疗中具有临床应用价值。
