摘要
目的:探究藏红花素对视网膜母细胞瘤自噬、凋亡、侵袭和PI3K/AKT活化的影响。方法:采用CCK-8法检测不同剂量藏红花素对视网膜母细胞瘤细胞Y79细胞活力的影响,选用0、1、2、4μmoL/L剂量藏红花素处理Y79细胞,分为藏红花素0μmoL/L组、藏红花素1μmoL/L组、藏红花素2μmoL/L组和藏红花素4μmoL/L组进行后续实验。免疫荧光检测LC3含量和AKT核转位;流式细胞仪检测细胞凋亡;Transwell检测细胞侵袭;免疫印迹检测LC3Ⅰ、LC3Ⅱ、Beclin1、ATG7、PI3K、p-PI3K、AKT、p-AKT蛋白表达水平。结果:与对照组相比较,藏红花素2、4μmoL/L组LC3含量显著升高,LC3Ⅱ/LC3Ⅰ比值显著升高,Beclin1、ATG7蛋白表达水平显著降低,细胞凋亡率显著升高,侵袭细胞数显著降低,p-PI3K/PI3K、p-AKT/AKT比值显著降低,AKT核转位显著降低。结论:藏红花素诱导视网膜母细胞瘤自噬性死亡,并抑制细胞侵袭和PI3K/AKT的活化。
Objective:To investigate the effects of Crocin on autophagy,apoptosis,invasion,and activation of PI3K/AKT in retinoblastoma.Methods:The effects of different doses of Crocin on the activity of retinoblastoma Y79 cells were detected by the CCK-8 method.Y79 cells treated with Crocin were divided into four groups:the Crocin 0 mol/L group,the Crocin 1μmol/L group,the Crocin 2μmol/L group,and the Crocin 4μmol/L group for subsequent experiments.LC3 content and AKT nuclear translocation were detected by immunofluorescence assay.Flow cytometry was used to detect apoptosis.Transwell was used to detected the cell invasion.Western blotting was used to detect the protein expression levels of LC3Ⅰ,LC3Ⅱ,Beclin1,ATG7,PI3K,p-PI3K,AKT and p-AKT.Results:Compared with the Crocin 0μmoL/L group,Crocin 2,4μmol/L group LC3 levels was significantly elevated,LC3Ⅱ/LC3Ⅰratio increased significantly,Beclin1,ATG7 protein expression level decreased significantly,the apoptosis rate increased significantly,invasive cells was significantly reduced,p-PI3K/PI3K,p-AKT/AKT ratio decreased significantly,nuclear translocation of AKT was significantly reduced.Conclusion:Crocin induces autophagic death of retinoblastoma and inhibits cell invasion and activation of PI3K/AKT.
作者
林珊
屈思萌
张荣
王生海
韦秋红
Lin Shan;Qu Simeng;Zhang Rong;Wang Shenghai;Wei Qiuhong(Department of Ophthalmology,Baoding First Central Hospital;ICU Department of CriticalCare Medicine,Affiliated Hospital of Hebei University,Baoding 071000,China)
出处
《解剖学杂志》
CAS
2022年第6期542-546,567,共6页
Chinese Journal of Anatomy
基金
保定市科技计划项目(17ZF260)。
