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ID:32832176
大小:2.57 MB
页数:42页
时间:2019-02-16
《nimesulide对低氧条件下人肝癌细胞hepg2的增殖抑制作用及机制探讨》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、南昌大学硕士学位论文Nimesulide对低氧条件下人肝癌细胞HepG2的增殖抑制作用及机制探讨姓名:吴梨华申请学位级别:硕士专业:肿瘤学指导教师:熊强201205摘要研究目的:探讨环氧合酶抑制剂Nimesulide对低氧条件下人肝癌细胞HepG2增殖的影响。研究内容和方法:应用Nimesulide处理1%02浓度条件下人肝癌细胞HepG2,分为6组:常氧(21%02)不加药组、常氧+501xmol/LNimesulide组、缺氧(1%02)不加药组、缺氧+50p,mol/LNimesulide组、缺氧+1001amol/LNimesulide组、缺氧+2001.tmol/
2、LNimesulide组,分别编号1-6组。采用四唑盐(MTT)法测HepG2细胞的增殖抑制率;RT-PCR测HepG2细胞中环氧合酶.2(COX.2)、HIF.1a和survivinmRNA表达的变化:Western印迹法测HepG2细胞中COX.2、HIF.1a和survivin蛋白表达的变化。结果:Nimesulide对低氧条件下HepG2细胞有增殖抑制的作用,呈时间.剂量依赖效应。与常氧比较,低氧条件下人HepG2细胞的COX.2、HIF.1a和survivin表达升高(尸3、RNA表达水平下调(P4、epG2cellsunderthehypoxiaenvironment.Contentsandmethodsofresearch:HepG2cellsweredividedinto6groups:normaxia(21%02)+cellswithnotreatment,nomaxia+cellswith50I.tmol/LNimesulide,hypoxia(1%02)+cellswithnotreatment,hypoxia+cellswith501xmol/LNimesulide,hypoxia+cellswith100pmol/LNimesulide,hypoxia+5、cellswith2001amol/LNimesulide.Thecellproliferationwasexaminedbymethylthiazolyltetrazolium(MTT)assay.Theexpressionlevelsofcyclooxygenase-2(cox一2),HIF-laandsurvivinmRNAweredetectedbyRT-PCR.TheCOX.2,HIF.1etandsurvivinproteinexpressionweremeasuredbyWesternblotting.Results:Nimesulideinhibitedth6、eproliferationoftheHepG2underthehypoxiaenvironment.Comparedwithcellsundernomarxiaenvironment,theexpressionofCOX-2.HIF.1仪andsurvivinoftheceilsunderhypoxiaenvironmentwerehigher(P<0.05).Comparedwiththehypoxia(1%02)+cellswithnotreatmentgroup,theexpressionlevelsofCOX一2andsurvivinmRNAofHepG2were7、obviouslowerinthetreatmentgroup(P
3、RNA表达水平下调(P4、epG2cellsunderthehypoxiaenvironment.Contentsandmethodsofresearch:HepG2cellsweredividedinto6groups:normaxia(21%02)+cellswithnotreatment,nomaxia+cellswith50I.tmol/LNimesulide,hypoxia(1%02)+cellswithnotreatment,hypoxia+cellswith501xmol/LNimesulide,hypoxia+cellswith100pmol/LNimesulide,hypoxia+5、cellswith2001amol/LNimesulide.Thecellproliferationwasexaminedbymethylthiazolyltetrazolium(MTT)assay.Theexpressionlevelsofcyclooxygenase-2(cox一2),HIF-laandsurvivinmRNAweredetectedbyRT-PCR.TheCOX.2,HIF.1etandsurvivinproteinexpressionweremeasuredbyWesternblotting.Results:Nimesulideinhibitedth6、eproliferationoftheHepG2underthehypoxiaenvironment.Comparedwithcellsundernomarxiaenvironment,theexpressionofCOX-2.HIF.1仪andsurvivinoftheceilsunderhypoxiaenvironmentwerehigher(P<0.05).Comparedwiththehypoxia(1%02)+cellswithnotreatmentgroup,theexpressionlevelsofCOX一2andsurvivinmRNAofHepG2were7、obviouslowerinthetreatmentgroup(P
4、epG2cellsunderthehypoxiaenvironment.Contentsandmethodsofresearch:HepG2cellsweredividedinto6groups:normaxia(21%02)+cellswithnotreatment,nomaxia+cellswith50I.tmol/LNimesulide,hypoxia(1%02)+cellswithnotreatment,hypoxia+cellswith501xmol/LNimesulide,hypoxia+cellswith100pmol/LNimesulide,hypoxia+
5、cellswith2001amol/LNimesulide.Thecellproliferationwasexaminedbymethylthiazolyltetrazolium(MTT)assay.Theexpressionlevelsofcyclooxygenase-2(cox一2),HIF-laandsurvivinmRNAweredetectedbyRT-PCR.TheCOX.2,HIF.1etandsurvivinproteinexpressionweremeasuredbyWesternblotting.Results:Nimesulideinhibitedth
6、eproliferationoftheHepG2underthehypoxiaenvironment.Comparedwithcellsundernomarxiaenvironment,theexpressionofCOX-2.HIF.1仪andsurvivinoftheceilsunderhypoxiaenvironmentwerehigher(P<0.05).Comparedwiththehypoxia(1%02)+cellswithnotreatmentgroup,theexpressionlevelsofCOX一2andsurvivinmRNAofHepG2were
7、obviouslowerinthetreatmentgroup(P
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