迷迭香酸通过调控PI3K通路诱导急性T细胞白血病Jurkat细胞自噬和细胞凋亡

Rosmarinic acid induce autophagy and apoptosis of T-cell acute leukemia cell line Jurkat via regulating PI3K pathway

目的:探究迷迭香酸(RA)对急性T细胞白血病Jurkat细胞存活的作用及机制。方法:将细胞随机分为Jurkat组、RA (5μmol/L)组、RA (10μmol/L)组和RA (20μmol/L)组,分别用0、5、10和20μmol/L的RA处理细胞,CCK8检测培养不同时间的细胞增殖倍数,克隆形成实验检测细胞增殖,流式检测细胞凋亡,免疫印迹检测Beclin1、P62、LC3、p-PI3K、PI3K、p-Akt、Akt和m TOR的表达,免疫荧光检测LC3的表达。结果:RA处理细胞4 d后,RA (5、10、20μmol/L)组细胞增殖倍数与Jurkat组比较明显降低,存活细胞数明显减少,细胞凋亡率明显升高;同时与Jurkat组比较,RA (5、10、20μmol/L)组细胞Beclin1表达水平和LC3Ⅱ/LCⅠ的比值明显降低,P62表达水平明显升高,LC3阳性表达与Jurkat组比较也明显减少;此外,RA (5、10、20μmol/L)还能显著降低p-PI3K/PI3K、p-Akt/Akt的比值和m TOR的表达水平。结论:RA可诱导急性T淋巴白血病Jurkat细胞自噬和细胞凋亡,作用机制可能与抑制PI3K/Akt通路激活有关。

Objective:To investigate the effects and mechanism of rosmarinic acid(RA)on the survival of T-cell acute leukemiacell line Jurkat.Methods:Jurkat cells were divided into Jurkat,RA(5μmol/L),RA(10μmol/L)and RA(20μmol/L)group and treated with RA at a concentrations of0,5,10and20μmol/L respectively.The cell proliferation rate was determined by CCK8assay,and cell survival was measured by colony formation assay.Flow cytometry was performed for cell apoptosis.The expression levels of Beclin1,p62,LC3,p-PI3K,PI3K,p-AKT,AKT and mTOR were measured by Western blot and immunofluorescence was employed to measure the positive expression of LC3.Results:Cell proliferation rate of RA(5,10,20μmol/L)group was decreased when compared with Jurkat group,cell survival was inhibited in RA(5,10,20μmol/L)group and cell apoptosis was induced by RA(5,10,20μmol/L).Meanwhile,compared with Jurkat group,the level of Beclin1and the ratio of LC3Ⅱ/LCⅠwere up-regulated,the expression level of P62was decreased significantly,and the positive expression of LC3was increased when compared with Jurkat group.In addition,RA(5,10,20μmol/L)down-regulated the ratio of p-PI3K/PI3K and p-Akt/Akt of Jurkat cells.Conclusion:RA induces autophagy and apoptosis of T-cell acute leukemia,and the machaism may be related to PI3K/Akt pathway.