摘要
目的:研究海参多糖对肾癌细胞自噬的影响。方法:将肾癌ACHN细胞分为海参多糖(50、100、200 mg/L)组和对照组,采用MTT法检测细胞活力,结晶紫染色观察细胞形态和细胞数量,Western blot法检测细胞增殖标志物(Ki-67、PCNA)、凋亡标志物(Cleaved Caspase-3、Cleaved Caspase-9)、自噬相关蛋白(Beclin1、P62、LC3-Ⅰ/LC3-Ⅱ)、AKT/mTOR通路相关蛋白(PI3K/p-PI3K、AKT/p-AKT、mTOR)的表达,FITC-V和PI染色法检测细胞凋亡率。结果:与对照组比较,海参多糖组细胞活力、球体内细胞数、Ki-67及PCNA表达降低,细胞凋亡率增加,Cleaved Caspase-3、Cleaved Caspase-9、Beclin1表达增加,P62表达减少,PI3K/p-PI3K、AKT/p-AKT、mTOR表达量减少(P<0.05);且随着海参多糖质量浓度的增加,上述作用逐渐增强。结论:海参多糖通过抑制AKT/mTOR信号通路进而抑制肾癌细胞增殖,并诱导肾癌细胞发生自噬。
Aim:To study the effects of sea cucumber polysaccharide on autophagy in renal cancer cells.Methods:ACHN cells were divided into sea cucumber polysaccharide(50,100,200 mg/L)groups and control group.Cell viability was detected by MTT method,and cell morphology and cell number were observed by crystal violet staining.Western blot was used to detect the expressions of the proliferation-associated proteins,the apoptosis-associated proteins,autophagy-associated proteins,the AKT/mTOR signaling pathway-associated proteins.Apoptosis was detected by FITC-V and PI staining.Results:Compared with the control group,the cell viability,the number of cells,and the expressions of Ki-67 and PCNA decreased,while the apoptosis rate increased in the sea cucumber polysaccharide groups(P<0 05).The expressions of Cleaved Caspase-3,Cleaved Caspase-9,Beclin1 increased,while the expression of P62,PI3K/p-PI3K,AKT/p-AKT,and mTOR decreased(P<0.05).The effects were dose-dependent.Conclusions:Sea cucumber polysaccharide could inhibit the proliferation and induce autophagy of renal cancer cells by inhibiting AKT/mTOR signaling pathway.
作者
李天
周东梅
刘一帆
江先汉
谢清灵
黄奕桥
殷羽飞
LI Tian;ZHOU Dongmei;LIU Yifan;JIANG Xianhan;XIE Qingling;HUANG Yiqiao;YIN Yufei(Department of Urology,the Fifth Affiliated Hospital,Guangzhou Medical University,Guangzhou 510700;Department of Traditional Chinese Medicine,the Fifth Affiliated Hospital,Guangzhou Medical University,Guangzhou 510700)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2019年第5期758-762,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
广东省中医药管理局基金项目(20171188)
