青蒿酯通过线粒体凋亡途径诱导Y79细胞凋亡

Artemisinate Induces Y79 Cell Apoptosis through Mitochondrial Ap­optosis Pathway

目的探究青蒿酯对视网膜母细胞瘤细胞的影响及机制.方法采用CCK-8法检测不同剂量青蒿酯对视网膜母细胞瘤细胞Y79细胞活力的影响,将细胞分为4组:Artesunate 0、30、60和120μmol/L组,分别选用0、30、60、120μmol/L青蒿酯处理Y79细胞.克隆形成实验检测细胞克隆形成率;流式细胞仪检测细胞凋亡率;流式分选检测线粒体膜电位;试剂盒检测谷胱甘肽(glutathione,GSG)、乳酸脱氢酶(lactate dehydrogenase,LDH)含量;蛋白免疫印迹检测PON2、Bax、Bcl-2蛋白表达水平.结果与Ar-tesunate 0μmol/L组相比较,Artesunate 60、120μmol/L组克隆形成率降低(P<0.05),细胞凋亡率降低(P<0.05),线粒体膜电位明显降低,GSH含量降低(P<0.05),LDH含量升高(P<0.05),PON2蛋白水平降低(P<0.05),Bax/Bcl-2比值升高(P<0.05).结论青蒿酯可能通过下调PON2表达介导线粒体凋亡途径诱导视网膜母细胞瘤细胞凋亡.

Objective To investigate the effect of artemisinate on the retinoblastoma cells and the mechanism.Methods CCK-8 method was used to detect the effect of artemisinate of different doses on the viability of retinoblastoma Y79 cells.Y79 cells were treated with artemisinate at 0,30,60 and 120μmol/L and were divided,into 4 groups in terms of the artemisinate doses.The rate of cell clone formation was determined by clonal formation assay.The apoptosis rate was flow cytomet-rically detected.Mitochondrial membrane potential was measured by flow sorting.The contents of GSH and LDH were detennined by using a specific kit.The protein expression levels of PON2,Bax and Bcl-2 were detected by Western blotting.Results Compared with artesunate 0μmol/L group,the clonal formation rate,apoptosis rate,mitochondrial membrane potential and GSH content in the artesunate 60 and 120μol/L groups were significantly lower(P<0.05).LDH content was increased(P<0.05),PON2 protein level decreased(P<0.05),and Bax/Bcl-2 ratio elevated(P<0.05).Conclusion Artemisinate might induce retinoblastoma cell apoptosis by down-regulating PON2 expression and mediating mitochondrial apoptosis.