基于Nrf2/ROS信号通路探讨岩藻多糖对食管癌细胞增殖的抑制作用  

Inhibition of Proliferation of Esophageal Cancer Cells by Fucoidan Based on Nrf2/ROS Signaling Pathway

作　　者：马永超[1] 程琦[1] 吴华[1] 陆琼[1] 金少举[1] MA Yongchao;CHENG Qi;WU Hua;LU Qiong;JIN Shaoju(Luohe Medical College,Luohe 462002,China)

机构地区：[1]漯河医学高等专科学校,河南漯河462002

出　　处：《食品工业科技》2024年第11期316-322,共7页Science and Technology of Food Industry

基　　金：河南省高等学校重点科研项目(22B310006);河南省自然科学基金项目(232300420068)。

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:1~16µg/mL岩藻多糖极显著抑制ECA109细胞增殖,48 h IC50为3.26µg/mL。与对照组(0.1%DMSO)相比,1、2、4µg/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。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 16µg/mL,and the IC50 was 3.26µg/mL at 48 h.Compared with the control group(0.1%DMSO),ECA109 cells treated with 1,2,and 4µg/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 

分 类 号：TS201.4[轻工技术与工程—食品科学]