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1、史塑王墨查(皇王生旦箜鲞箜塑墅d1StemCell(ElectronicEdition),May2015,7o1.5,No.243·論著·染料木素抑制人胰腺癌細(xì)胞株P(guān)ANC.1細(xì)胞增殖機(jī)制的研究馬志亮崔嘯晨李夏青王麗鳳苑洪蓮衛(wèi)建平安峽尹彩虹王志茹【摘要】目的探討不同濃度染料木素對胰腺癌細(xì)胞作用及其抑癌基因表達(dá)的研究。方法本實(shí)驗(yàn)共分實(shí)驗(yàn)組(80,120,160,200gmol/L)的染料木素、空白對照組(PBS)和標(biāo)準(zhǔn)對照(吉西他濱)。各組培養(yǎng)基處理細(xì)胞12,24,36,48,60h后,細(xì)胞計(jì)數(shù)試劑盒(CC
2、K.8)法檢測人胰腺癌細(xì)胞株P(guān)ANC.1在染料木素作用下的增殖活性;流式細(xì)胞術(shù)檢測人胰腺癌細(xì)胞株P(guān)ANC.1在染料木素木素作用下的凋亡細(xì)胞百分率;用逆轉(zhuǎn)錄一聚合酶連反應(yīng)(RT_pCR)方法測定P53及P21抑癌基因的表達(dá);Westem.blot法檢測抑癌基因P53和P21的蛋白表達(dá)水平。細(xì)胞凋亡與細(xì)胞基因mRNA采用的是LSD法,細(xì)胞增殖數(shù)據(jù)采用的是配對t檢驗(yàn)。結(jié)果流式細(xì)胞術(shù)顯示,人胰腺癌細(xì)胞株P(guān)ANC一1處理后60h時(shí)在不同濃度(80,120,160,200gmol/L)的染料木素作用下細(xì)胞凋亡率分別為
3、(3.724-1.58)%、(35.984-3.76)%、(51.364-4.32)%和(64.104-2.09)%。吉西他濱(gemcitabine,GEM)組為(43.454-5.28)%,不同濃度染料木素組與標(biāo)準(zhǔn)對照組間比較差異有統(tǒng)計(jì)學(xué)意義(均P<0.O1);細(xì)胞增殖效果:160~tmol/L與200pmol/L、120~tmol/L與200~tmol/L、200gmol/L與吉西他濱組之間比較,均有統(tǒng)計(jì)學(xué)意義(P<0.01),而160pmol/L與吉西他濱組比較無統(tǒng)計(jì)學(xué)意義(P>0.05);Wes
4、tern.blot檢測蛋白水平顯示160gmol/L和200gmol/L染料木素相對于空白對照能夠明顯的增強(qiáng)抑癌基因P21和P53的蛋白水平的表達(dá),相對于標(biāo)準(zhǔn)組沒有明顯的差異。結(jié)論染料木素可以抑制人胰腺癌細(xì)胞株P(guān)ANC—l增殖,并且初步探索了這種抑制作用表現(xiàn)出濃度及時(shí)間依賴性。【關(guān)鍵詞】胰腺腫瘤;染料木素;吉西他濱;細(xì)胞凋亡GenisteininhibitsproliferationofPANC-1humanpancreaticcancercellsMaZhiliang~,CuiXiaochen,LiXi
5、aqing2,WangLifeng3,YuanHonglian4,WeiJianping5,AnXia;HnCaihong6WangZhiru.DepartmentofGeneralSurgery,OperationRoom,~DepartmentofPathology,SecondHospitalofShanxiMedicalUniversity,Tab,uan030001China;2Departmentofpathophysiology,ShanxiMedicalUniversity,Taiyuan
6、030001,China;3DepartmentofPathology,ShanxiDayihospital,Ta&uan030001,China;~DepartmentofInternalMedicine,ShuozhouCentralHospital,Shuozhou03600~ChinaCorrespondingauthor."WangZhiru,Email:geraldzwang@yahoo.corn【Abstract】ObjectiveToexploretheeffectsofGenistein
7、onpancreaticcancerandtheexpressionoftumorsuppressorgenes.MethodsThisstudyincludedexperimentalgroup(80,120,160,200pmol/LGenistein),positivecontrolgroup(gemcitabine)andblankgroup(PBS).Atthedifferenttimepoints(12,24,36,48and60h)proliferationofhumanpancreatic
8、cancercelllinePANC一1wasevaluatedwithCCK一8assay.ApoptoticcellsweredetectedwithFCM.ExpressoftumorsuppressorgenesP53andP21wasevaluatedusingDOh10.3877/cmad.issn.2095ol221.2015.02.009基金項(xiàng)目:山西省歸國留學(xué)項(xiàng)目(2010099)作者單位:030001太原,