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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

Crocin induces autophagic death of retinoblastoma and inhibits cell invasion and activation of PI3K/AKT

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【作者】 林珊屈思萌张荣王生海韦秋红

【Author】 Lin Shan;Qu Simeng;Zhang Rong;Wang Shenghai;Wei Qiuhong;Department of Ophthalmology, Baoding First Central Hospital;ICU Department of Critical Care Medicine, Affiliated Hospital of Hebei University;

【通讯作者】 韦秋红;

【机构】 保定市第一中心医院眼一科河北大学附属医院ICU重症医学科

【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。

【Abstract】 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 1 μmol/L group, the Crocin 2 μmol/L group, and the Crocin 4 μmol/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 0 μmoL/L group, Crocin 2, 4 μmol/L group LC3 levels was significantly elevated, LC3Ⅱ/LC3Ⅰ ratio 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.

【基金】 保定市科技计划项目(17ZF260)
  • 【文献出处】 解剖学杂志 ,Chinese Journal of Anatomy , 编辑部邮箱 ,2022年06期
  • 【分类号】R285
  • 【下载频次】15
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