p38fasfasl在罗格列酮诱导人胃癌sgc7901细胞凋亡中调控作用word论文.docxVIP

P38/Fas/Fasl在罗格列酮诱导人胃癌SGC7901 细胞凋亡中的调控作用硕士研究生：朱梦霞导师：张琍教授中文摘要目的：研究过氧化物酶体增殖因子活化受体(Peroxisomeproliferator- activated receptor gamma, PPARγ)的重要配体罗格列酮（Rosiglitazone, ROS）对人胃低分化腺癌细胞株SGC7901凋亡的影响及p38/Fas/Fasl途径是否能调控胃癌细胞的凋亡。方法：首先于体外培养人胃癌SGC7901细胞至对数生长期，再分别用不同的浓度的ROS处理SGC7901细胞，48小时之后，采用流式细胞术（Flow cytometrymethod, FCM）检测细胞凋亡；RT-PCR法观察细胞p38,Fas ,Fasl mRNA的表达；Western Blot 法测定p-p38, Fas, Fasl 蛋白的表达。结果：ROS能促进胃癌细胞株SGC7901的凋亡，且呈浓度依赖效应。流式细胞术检测显示ROS（25μmol/l、50μmol/l、100μmol/l）分别作用于SGC7901 细胞48h后，三组细胞的凋亡率分别为26.67％±4.50％、53.67％±3.51％、72.3 3％±3.06％，与溶媒对照组相比，细胞凋亡率显著增强（P0.05）。不同浓度的罗格列酮作用后能显著增强SGC7901细胞的p38, Fas, Fasl mRNA和p-p38, Fas,Fasl蛋白的表达（P0.05）。结论：ROS能呈浓度依赖性的诱导胃癌SGC7901细胞的凋亡，其诱导细胞凋亡的机制可能是通过促进p38的表达和（或）p38的磷酸化，从而上调F as/Fasl 的表达，最终诱导SGC7901 细胞凋亡。关键词：罗格列酮；凋亡；p38MAPK;Fas; Fasl2Theregulatoryroleofp38/Fas/Faslon the rosiglitazone-induced apoptosisin humangastric cancer celllineMGC7901ABSTRACTObjectives:Toinvestigatetheeffectsofperoxisomeproliferator-activated receptorgamma(PPARγ)ligandrosiglitazone(ROS)onapoptosisinhumangastric lower-differentiationadenocarcinomacelllineSGC7901,andtheregulatoryroleof p38/Fas/Fasl on the rosiglitazone-induced apoptosis in MGC7901.Methods:First,gastricSGC7901cellswereculturedinvitro,cellsof exponentialphaseofgrowthareusedtoexperiment.Usingdifferentconcentrationsof ROSroleinSGC7901,48hourslater,flowcytometrymethodwasusedtodetectthe cellapoptosis,theexpressionofmRNAofp38,Fas,FaslwastestedbyRT-PCR,the expression ofprotein ofp-p38, Fas,Fasl was checked byWestern-Blot assay.Results:ROScouldpromotetheapoptosisofgastriccelllineSGC7901ina dose-dependentmanner.Flowcytometrymethodshowedthat,ROS(25μmol/l, 50μmol/l,100μmol/l)roleinSGC7901for48hours,theapoptosisratesofthethreegroupswere26.67％±4.50％,53.67％±3.51％,72.33％±3.06％,theapoptosisoftheexperimentalgroupsweresignificantlyhigherthanthecontrolgroup(P0.05). DifferentconcentrationsofROSpromotedtheexpressionofmRNAofp38,Fas,Fasl and proteinofp-p38,Fas, Fasl in SGC7901 cells(P0.05).Conclusions:ROScanpromotetheapoptosisof