12-脂氧化酶影响胃癌细胞AGS中P21及bcl-2的表达  

12-Lipoxygenase Affect the Expression of bcl-2 and P21 in Human Gastric Cancer AGS Cells

作　　者：陈丰霖[1] 王小众[1] 李建英[1] 陈治新[1] 黄月红[1] 

机构地区：[1]福建医科大学附属协和医院消化科,福州350001

出　　处：《福建医科大学学报》2011年第5期323-326,共4页Journal of Fujian Medical University

基　　金：福建省科技厅科研基金(2007F3034)

摘　　要：目的研究12-脂氧化酶(12-LOX)对胃癌细胞中P21、bcl-2及磷酸化ERK1/2(p-ERK1/2)表达的影响及与ERK1/2信号传导通路的关系。方法胃癌细胞AGS常规培养于含10%胎牛血清的RPMI-1640培养液中24h至对数生长期,改用无血清RPMI-1640培养基培养24h加入干预药物。P21、bcl-2及p-ERK1/2表达采用Western blot法测定。分别采用100nmol/L的12-LOX催化合成产物12-羟基廿碳四烯酸(12-HETE)及40μmol/L的12-LOX特异性抑制黄苓素干预AGS细胞24h,测定P21、bcl-2及p-ERK1/2含量;采用ERK抑制剂PD98059预处理AGS细胞2h后,再分别用100nmol/L的12-HETE及40μmol/L黄苓素干预24h后再测定P21、bcl-2及p-ERK1/2含量。结果经100nmol/L的12-HETE干预24h后,p-ERK1/2、P21、bcl-2均显著高于末干预组(P<0.05),而40μmol/L黄苓素干预24h后,p-ERK1/2、P21、bcl-2显著低于对照组(P<0.05);采用ERK抑制剂PD98059预处理AGS细胞2h后再分别用100nmol/L的12-HETE及40μmol/L黄苓素干预与未采用PD98059预处理组相比,PD98059预处理后12-HETE干预组bcl-2水平低于仅用12-HETE干预组,PD98059预处理后黄苓素干预组bcl-2水平高于仅用黄苓素干预组(P<0.05)。而PD98059预处理后2组P21水平含量均无明显差异。结论 12-LOX对胃癌细胞P21及bcl-2表达具有调节作用,12-LOX通过其合成产物12-HETE促进ERK1/2的磷酸化,并通过ERK1/2途径调节bcl-2的表达,P21的表达不受ERK1/2途径调控。Objective To investigate the effects of 12-lipoxygenase（12-LOX） and its inhibitor,baicalein,on ERK1/2 activation and the expression of bcl-2 and P21 in human gastric cancer AGS cells.Methods Human gastric cancer AGS cells were cultured in a RPMI-1640 medium with 10% fetal bovine serum（FBS）.After overnight incubation,AGS cells were cultured in a fresh RPMI1640 medium without FBS for another 24 h and then treated with various drugs.AGS cells were treated with 12-LOX inhibitor,baicalein,or its metabolite 12-HETE respectively.The concentration of phosphorylated ERK（p-ERK）,bcl-2 and P21 was examined by Western-blot.Results Western blotting analysis results revealed that the concentration of p-ERK1/2 in cells treated with 100 nmol/L 12-HETE was significantly（P0.05） higher than in the untreated group and the concentration of phosphorylated ERK1/2 in cells treated with 40 mol/L baicalein was significantly（P0.05） lower than in the untreated group.The expression level of bcl-2 and P21 was up-regulated and down-regulated after separate treatment with 12-HETE and baicalein respectively,and these effects on the expression of bcl-2 could be blocked by PD98059.Conclusions 12-LOX activates ERK1/2 pathway,12-LOX affect the expression of P21 and bcl-2.

关 键 词：胃肿瘤 脂氧化酶 花生四烯酸类 原癌基因蛋白质C-BCL-2 原癌基因蛋白质p21(ras) 肿瘤细胞 培养的 

分 类 号：R735.2[医药卫生—肿瘤]