丙烯酰胺对SH-SY5Y神经细胞凋亡和miR-21表达的影响

Effects of acrylamide on apoptosis and miR-21 expression in SH-SY5Y cells

目的:探讨丙烯酰胺(AA)对人SH-SY5Y神经细胞凋亡和miR-21表达的影响。方法:不同浓度丙烯酰胺作用于人神经母细胞瘤SH-SY5Y细胞24 h后,采用四甲基偶氮唑盐(MTT)法测定丙烯酰胺对SH-SY5Y细胞活力的影响,Hoechst 33258荧光染色法检测细胞凋亡,real-time PCR检测miR-21表达水平,Western blot检测PTEN、p-AKT、AKT、Bcl-2、Bax、caspase 9和caspase 3蛋白表达。结果:丙烯酰胺剂量依赖性地降低SH-SY5Y细胞活性,Hoechst 33258染色显示明显细胞凋亡形态变化;丙烯酰胺显著降低SH-SY5Y细胞miR-21的表达,同时升高PTEN、Bax、caspase 9、caspase 3水平并降低p-AKT和Bcl-2表达。结论:丙烯酰胺可通过线粒体途径诱导SH-SY5Y神经细胞凋亡,其机制可能与丙烯酰胺引起miR-21表达下调有关。

Objective：To investigate the effects of acrylamide on apoptosis and miR-21 expression in SH-SY5Y neuroblastoma cells.Methods：Cell viability of SH-SY5Y cells treated with different concentrations of acrylamide for 24 hours was measured by MTT.Hoechst 33258 staining was used for the determination of cell apoptosis;miR-21 expression was detected by real-time PCR;the expression levels of PTEN,p-AKT,AKT,Bcl-2,Bax,caspase 9 and caspase 3 were examined by Western blot.Results：Acrylamide exposure resulted in decreased viability and induction of apoptosis of SH-SY5Y cells in a dose-dependent manner.miR-21 expression was downregulated by acrylamide.Meanwhile,acrylamide increased the expression levels of PTEN,Bax,caspase 9 and caspase 3,and decreased p-AKT and Bcl-2 expression in SH-SY5Y cells.Conclusion：Acrylamide induces apoptosis of SH-SY5Y cells via mitochondrion pathway,and down-regulation of miR-21 may be involved in the regulation of apoptotic process induced by acrylamde.