Inhibition of Proliferation of Esophageal Cancer Cells by Fucoidan Based on Nrf2/ROS Signaling Pathway
马永超 1程琦 1吴华 1陆琼 1金少举1
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作者信息
1. 漯河医学高等专科学校,河南漯河 462002
折叠
摘要
目的:探讨岩藻多糖对食管癌增殖的影响,并分析其机制.方法:MTT法分析细胞增殖抑制率,Hoechst 33258染色和流式细胞术检测细胞凋亡,DCFH-DA探针检测ROS水平,Western blot法分析Nrf2、HO-1、NQO-1、Bcl-2、Bax、caspase-3水平,研究岩藻多糖对细胞增殖以及Nrf2/ROS信号通路的影响.裸鼠成瘤实验验证岩藻多糖对瘤体的瘤重、瘤体积及Nrf2、HO-1、NQO-1水平的影响.结果:1～16 μg/mL岩藻多糖极显著抑制ECA109细胞增殖,48 h IC50为3.26 μg/mL.与对照组(0.1％DMSO)相比,1、2、4 μg/mL岩藻多糖处理后的ECA109细胞出现核凝聚、染色质不规则收缩、凋亡小体等明显的凋亡特征,(极)显著促进ECA109细胞凋亡,极显著下调Bcl-2表达水平,极显著上调Bax、Cleaved-caspase-3表达水平,极显著增加ROS水平,极显著降低Nrf2、HO-1、NQO-1蛋白水平(P＜0.05,P＜0.01);Nrf2过表达能显著下调岩藻多糖抑制ECA109细胞增殖效果,显著下调ROS水平,显著上调Nrf2、HO-1、NQO-1蛋白水平(P＜0.05).体内实验显示,50、100 mg/kg岩藻多糖极显著抑制瘤体体积、瘤体质量,下调瘤体Nrf2、HO-1、NQO-1水平(P＜0.05).结论:岩藻多糖抑制ECA109细胞增殖,对体内移植瘤抑瘤效果显著,其机制与调控Nrf2/ROS信号通路有关.
Abstract
Objective:To explore the effect of fucoidan on esophageal cancer and analyze its mechanism.Methods:MTT assay was used to analyze the inhibition rate of cell proliferation,Hoechst33258 staining and flow cytometry were used to detect cell apoptosis,and DCFH-DA probe was used to detect ROS level,and Western blot was used to analyze levels of Nrf2,HO-1,NQO-1,Bcl-2,Bax and caspase-3,which were used to observe its effects on fucoidan-regulated cell proliferation and Nrf2/ROS signaling pathway.The tumor formation experiment in nude mice verified the effects of fucoidan on tumor weight,tumor volume and levels of Nrf2,HO-1 and NQO-1 in nude mice.Results:The proliferation of ECA109 cells was significantly inhibited by fucoidan from 1 to 16 μg/mL,and the IC50 was 3.26 μg/mL at 48 h.Compared with the control group(0.1％DMSO),ECA109 cells treated with 1,2,and 4 μg/mL fucoidan showed obvious apoptotic characteristics,such as nuclear condensation,irregular chromatin contraction and apoptotic bodies,which(extremely)significantly promoted the apoptosis of ECA109 cells and significantly down-regulated the expression level of Bcl-2.The expression levels of Bax and Cleaved-caspase-3 were significantly increased,ROS levels were significantly increased,and Nrf2,HO-1,and NQO-1 protein levels were significantly decreased(P＜0.05,P＜0.01).Nrf2 overexpression could significantly down-regulate the inhibitory effect of fucoidan on ECA109 cell proliferation,significantly down-regulate ROS levels,and significantly up-regulate Nrf2,HO-1,NQO-1 protein levels(P＜0.05).In vivo experiments showed that 50 mg/kg and 100 mg/kg of fucoidan significantly inhibited tumor volume and mass,and down-regulated the levels of Nrf2,HO-1 and NQO-1 in tumors(P＜0.05).Conclusion:Fucoidan inhibits the proliferation of ECA109 cells and has significant anti-tumor effect on transplanted tumor in vivo,and its mechanism is related to the regulation of Nrf2/ROS signal pathway.
关键词
岩藻多糖/Nrf2/ROS信号通路/食管癌/细胞增殖/移植瘤
Key words
fucoidan/Nrf2/ROS signal pathway/esophageal cancer/cell proliferation/transplanted tumor
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