College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China.
Department of Pharmaceuticals, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China.
Phytomedicine. 2016 Mar 15;23(3):274-82. doi: 10.1016/j.phymed.2016.01.012. Epub 2016 Feb 8.
We previously reported the effect of dioscin on human gastric carcinoma SGC-7901 cells, but its effects on other gastric cancers are still unknown.
The present paper aimed to demonstrate the activity of dioscin against human gastric carcinoma MGC-803 and MKN-45.
In our study, MGC-803 and MKN-45 cells were used to examine the effects of dioscin on human gastric carcinoma in vitro. The effects of dioscin against human gastric carcinoma in vivo were accomplished by the xenografts of MGC-803 cells in BALB/c nude mice.
AO/EB and DAPI staining, TEM, single cell gel electrophoresis and flow cytometry assays were used in cell experiments. Then, an iTRAQ-based proteomics approach, DNA and siRNA transfection experiments were carried out for mechanism investigation.
In MGC-803 cells, dioscin caused DNA damage and mitochondrial change, induced ROS generation, Ca(2+) release and cell apoptosis, and blocked cell cycle at S phase. In vivo results showed that dioscin significantly suppressed the tumor growth of MGC-803 cell xenografts in nude mice. In addition, dioscin markedly inhibited cell migration, caused Cytochrome c release and adjusted mitochondrial signal pathway. Then, an iTRAQ-based proteomics approach was carried out and 121 differentially expressed proteins were found, in which five biomarkers associated with cell cycle, apoptosis and migration were evaluated. Dioscin significantly up-regulated the levels of GALR-2 and RBM-3, and down-regulated CAP-1, Tribbles-2 and CliC-3. Furthermore, overexpressed DNA transfection of CAP-1 enhanced cell migration and invasion, which was decreased by dioscin. SiRNA to Tribbles-2 affected the protein levels of Bcl-2, Bax and MAPKs, suggesting that dioscin decreased Tribbles-2 level leading to cell apoptosis.
Our works confirmed the activity of dioscin against gastric cancer. In addition, this work also provided that dioscin is a new potent candidate for treating gastric cancer in the future.
我们之前报道了薯蓣皂甙对人胃癌 SGC-7901 细胞的作用，但它对其他胃癌的作用尚不清楚。
本研究旨在探讨薯蓣皂甙对人胃癌 MGC-803 和 MKN-45 的作用。
本研究采用 MGC-803 和 MKN-45 细胞，体外观察薯蓣皂甙对人胃癌的作用。采用 MGC-803 细胞裸鼠异种移植模型，观察薯蓣皂甙体内对人胃癌的作用。
采用 AO/EB 和 DAPI 染色、透射电镜、单细胞凝胶电泳和流式细胞术进行细胞实验。然后，采用 iTRAQ 蛋白质组学方法、DNA 和 siRNA 转染实验进行机制研究。
在 MGC-803 细胞中，薯蓣皂甙引起 DNA 损伤和线粒体变化，诱导 ROS 生成、Ca(2+)释放和细胞凋亡，阻断细胞周期于 S 期。体内实验结果表明，薯蓣皂甙显著抑制裸鼠 MGC-803 细胞异种移植瘤的生长。此外，薯蓣皂甙明显抑制细胞迁移，引起细胞色素 c 释放，并调节线粒体信号通路。然后，采用 iTRAQ 蛋白质组学方法，发现 121 个差异表达蛋白，其中 5 个与细胞周期、凋亡和迁移相关的生物标志物进行了评估。薯蓣皂甙显著上调 GALR-2 和 RBM-3 的水平，下调 CAP-1、Tribbles-2 和 CliC-3。此外，CAP-1 的过表达 DNA 转染增强了细胞迁移和侵袭，而薯蓣皂甙则降低了细胞迁移和侵袭。用 Tribbles-2 的 siRNA 处理影响了 Bcl-2、Bax 和 MAPKs 的蛋白水平，表明薯蓣皂甙降低 Tribbles-2 水平导致细胞凋亡。
本研究证实了薯蓣皂甙对胃癌的活性。此外，本研究还表明薯蓣皂甙是未来治疗胃癌的一种新的有效候选药物。
