蛇床子素抑制PI3K/AKT/mTOR的活化诱导视网膜母细胞瘤Y79细胞凋亡和抑制裸鼠成瘤的研究

Effect of Osthole in Inducing Apoptosis of Retinoblastoma Y79 Cells and Inhibiting Tumor Formation in Nude Mice by Inhibiting Activation of PI3K/AKT/mTOR

目的研究蛇床子素通过抑制PI3K/AKT/mTOR的活化,诱导视网膜母细胞瘤Y79细胞凋亡和抑制裸鼠成瘤的作用。方法将Y79细胞分为对照组和蛇床子素8、16、32μmol/L组,对照组给予生理盐水,不同浓度蛇床子素组细胞分别给予蛇床子素8、16、32μmol/L干预。检测不同浓度蛇床子素对Y79细胞凋亡和胞内磷脂酰肌醇激酶(PI3K)、蛋白激酶B(AKT)、雷帕霉素靶蛋白(mTOR)mRNA及蛋白的影响。构建视网膜母细胞瘤裸鼠腋下异位移植瘤模型,并观察蛇床子素20、50、100 mg/kg处理的裸鼠成瘤情况。结果与对照组比较,8、16、32μmol/L的蛇床子素组细胞凋亡率均显著增加,且呈浓度依赖性(P<0.05)。与对照组比较,8、16、32μmol/L的蛇床子素凋亡蛋白Caspase-3、Caspase-9的mRNA及蛋白表达水平增高,PI3K、AKT、mTOR磷酸化水平减低,且呈浓度依赖性(P<0.05)。与对照组比较,20、50、100 mg/kg的蛇床子素组裸鼠视网膜母细胞瘤体积和重量降低,且呈剂量依赖性(P<0.05)。结论蛇床子素可通过抑制PI3K/AKT/mTOR的活化来诱导视网膜母细胞瘤Y79细胞凋亡和抑制裸鼠成瘤。

Objective To explore the effect of Osthole on inducing apoptosis of retinoblastoma Y79 cells and inhibiting tumor formation in nude mice by inhibiting activation of PI3K/AKT/mTOR.Methods Y79 cells were divided into control group and 8,16,32μmol/L Osthol groups.The control group was given normal saline,and groups with different concentrations of Osthol were given 8,16 and 32μmol/L Osthol for intervention respectively.The effects of different concentrations of Osthol on the apoptosis of Y79 cells and on the mRNA and protein of phosphatidylinositol kinase(PI3K),protein kinase B(AKT),rapamycin target protein(mTOR)in Y79 cells were detected.The heterotopic xenograft model of retinoblastoma in nude mice was established and the tumor formation in nude mice treated with 20,50,100 mg/kg Osthol respectively was observed.Results Compared with the control group,the apoptosis rate of 8,16 and 32μmol/L Osthol groups was increased significantly in a concentration-dependent manner(P<0.05).Compared with the control group,the mRNA and protein expression level of Caspase-3 and Caspase-9 in 8,16 and 32μmol/L Osthol groups were increased,and the phosphorylation level of PI3K,Akt and mTOR was significantly decreased in a concentration-dependent manner(P<0.05).Compared with the control group,the volume and weight of retinoblastoma in the 20,50,100 mg/kg Osthol group were decreased in a dose-dependent manner(P<0.05).Conclusion Osthole can induce apoptosis of retinoblastoma Y79 cells and inhibit tumor formation in nude mice by inhibiting activation of PI3K/Akt/mTOR.