p-ERK1/2在一氧化氮诱导的HepG2细胞凋亡中的作用  

Effects of p-ERK1/2 on nitric oxide donor induced apoptosis of HepG2 cells

作　　者：刘智勇[1] 牛志远[1] 郑薇[1] 沈萍萍[1] 

机构地区：[1]南京大学生命科学学院,医药生物技术国家重点实验室,南京210093

出　　处：《中国药科大学学报》2012年第6期530-534,共5页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.31070764)~~

摘　　要：探讨p-ERK1/2在一氧化氮诱导的人肝癌细胞HepG2凋亡中的作用。用不同浓度的硝普钠(sodium nitro-prusside,SNP)处理HepG2细胞12 h,通过运用荧光探针技术与Griess法检测细胞内一氧化氮浓度,流式细胞术测定肿瘤细胞的凋亡率,Western blot检测p-ERK1/2蛋白的表达水平。结果发现:一氧化氮通过上调p-ERK1/2的表达水平诱导HepG2细胞的凋亡,1 mmol/L SNP可以诱导HepG2细胞凋亡,胞内一氧化氮含量与p-ERK1/2蛋白表达量均高于对照组,转染ERK1过磷酸化质粒则明显促进肿瘤细胞的凋亡。实验结果显示,一氧化氮通过上调p-ERK1/2的表达水平诱导HepG2细胞的凋亡,此研究发现了p-ERK1/2在一氧化氮诱导肿瘤细胞凋亡过程中的新作用,可进一步深入探讨其相关机制。The role of p-ERK1/2 in nitric oxide induced cell apoptosis was investigated in this paper.Human hepatoma cells HepG2 were treated by sodium nitroprusside（SNP） in a dose-dependent manner with a control group.The amount of nitric oxide produced by SNP was determined by NO-DAF technique and Griess assay,respectively.Apoptosis of HepG2 cells was detected by flow cytometry and the level of p-ERK1/2 expression was determined by Western blot assay.Mutated ERK1 plasmid was constructed according to the handbook of site-directed gene mutagenesis kit and the plasmids were transfected by Lipofectamine 2000 reagent.Results showed 1 mmol/L SNP could effectively induce HepG2 cell apoptosis with increasing p-ERK1/2 expression and nitric oxide concentrations compared with the control group.Furthermore,pre-treatment of NO scavenger hemoglobin（HB） notably reduced the concentrations of nitric oxide and inhibited SNP induced cell apoptosis and activation of ERK1/2.Overexpression of dominate positive ERK1 plasmid which can phosphorylate PPARγ increased SNP induced cell apoptosis.These results suggest nitric oxide could induce cell apoptosis by up regulating the expression of p-ERK1/2,which implies the important role of p-ERK1/2 in nitric oxide induced cell apoptosis,and may shed a new light on the discovery of relevant mechanisms.

关 键 词：p-ERK1 2 一氧化氮 人肝癌细胞 HEPG2 细胞凋亡 

分 类 号：R965[医药卫生—药理学]