摘要
目的:基于蛋白激酶R样内质网激酶(PERK)-活化转录因子4(ATF4)-转录因子C/EBP同源蛋白(CHOP)信号通路探讨紫檀芪抑制结肠癌细胞增殖、迁移,诱导细胞凋亡,从而探究其抗结肠癌的作用。方法:通过细胞计数的盒子-8(CCK8)法、细胞划痕实验、流式细胞术等方法检测紫檀芪(PTE)作用于HCT116细胞后观察其对HCT116细胞增殖、迁移、凋亡和细胞周期的影响,通过活性氧(ROS)探针检测细胞内ROS的水平,并将HCT116细胞分为对照组、PTE组、PTE+GSK2606414(特异性PERK通路抑制剂)组,用蛋白质印迹法(Western blot)检测PERK-ATF4-CHOP信号通路相关蛋白PERK、p-PERK、ATF4、CHOP和凋亡相关蛋白BAX、BCL-2的表达。结果:PTE抑制HCT116细胞增殖,呈浓度依赖特性,其半抑制浓度(IC50)为114.8μmol·L^(-1);PTE可以显著抑制HCT116细胞的增殖和迁移,增加细胞的早期凋亡比率,诱导HCT116细胞的细胞周期阻滞在G1期,并且可诱导HCT116细胞ROS的水平过量增加;HCT116细胞内促凋亡蛋白BAX表达增加,抑凋亡蛋白BCL-2表达降低,PERK-ATF4-CHOP信号通路相关蛋白的表达增加;加入GSK2606414可逆转PTE对HCT116细胞的促凋亡作用。结论:PTE可显著抑制结肠癌细胞HCT116的增殖、迁移并诱导其发生细胞周期阻滞和凋亡;PTE诱导结肠癌细胞HCT116的凋亡过程,可能与激活PERK-ATF4-CHOP信号通路有关。
Objective Based on the protein kinase R-like endoplasmic reticulum kinase(PERK)-activated transcription factor 4(ATF4)-transcription factor C/EBP homologous protein(CHOP)signaling pathway,this study aims to investigate the inhibitory effect of purple sandalwood on colon cancer cell proliferation and migration,induce cell apoptosis,and exert its anti colon cancer effect.Methods The effects of pterostilbene(PTE)on the proliferation,migration,apoptosis and cell cycle of HCT116 cells were observed by means of cell counting kit-8(CCK-8)assay,cell scratch assay,flow cytometry and other methods.The levels of intracellular reactive oxygen species(ROS)were detected using ROS probes,and HCT116 cells were divided into control group,PTE group,and PTE+GSK2606414(specific PERK pathway inhibitor)group.Western blot was used to detect the expression of PERK-ATF4-CHOP signaling pathway related proteins PERK,p-PERK,ATF4,CHOP,and apoptosis related proteins BAX and BCL-2.Results PTE inhibited the proliferation of HCT116 cells in a concentration dependent manner,with a half maximal inhibitory concentration(IC50)of 114.8μmol·L^(-1);PTE can significantly inhibit the proliferation and migration of HCT116 cells,increase the early apoptosis rate of cells,induce cell cycle arrest in G1 phase of HCT116 cells,and induce excessive increase in ROS levels in HCT116 cells;The expression of pro apoptotic protein BAX increased and the expression of anti apoptotic protein BCL-2 decreased in HCT116 cells;The expression of PERK-ATF4-CHOP signaling pathway related proteins increased;Adding GSK2606414 can reverse the pro apoptotic effect of PTE on HCT116 cells.Conclusion PTE can significantly inhibit the proliferation,migration,and induce cell cycle arrest and apoptosis of colon cancer cell line HCT116;The induction of apoptosis in colon cancer cell line HCT116 by PTE may be related to the activation of the PERK-ATF4-CHOP signaling pathway.
作者
王雅鑫
郑晓悦
付文亮
WANG Yaxin;ZHENG Xiaoyue;FU Wenliang(Department of Human Anatomy,School of Basic Medical Sciences,Chengde Medical College,Hebei Chengde 067000)
出处
《深圳中西医结合杂志》
2025年第10期5-9,I0004,I0005,I0006,I0007,共9页
Shenzhen Journal of Integrated Traditional Chinese and Western Medicine
基金
承德医学院校级科研课题(201904)
承德医学院人体解剖与组织胚胎学优势学科资助项目(承医政发[2023]22号)。
关键词
紫檀芪
结肠癌
内质网应激
凋亡
Pterostilbene
Colon cancer
Endoplasmic reticulum stress
Apoptosis
