刺参酸性粘多糖对肝肿瘤hepg2细胞增殖及相关基因表达影响

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1、及作用于细胞周期G1期的Rb—E2F通路，抑制其E2F一1、cyclinDl、CDK4基因及蛋白的表达，促进pRb和P21基因及蛋白的表达，使HepG2细胞发生G1期阻滞。同时，SJAMP对正常肝细胞张氏细胞的生长没有影响。硕士研究生：金岩(营养与食品卫生学)指导教师：宋扬教授关键词：刺参酸性粘多糖；HepG2细胞；张氏细胞；细胞增殖AbstractObjective：ToobservetheeffectofStichopusJaponicusacidmucopolysaccharide(SJAMP)on

2、HepG2cellsproliferation，then,tostudytheeffectandmechanismofSJAMPonproliferationofHepG2cells，toprovidethebasisfortheapplicationofSJAMP．Methods：livercancercellsHepG2wereculturedinDMEMmedium(highglucose)，withstableOutammesupplementedwith10％FBSandincubatedat3

3、7。Cwith5％C02．TheculturemediumoflivercellsChangWaSPRMI1640．WhenthecellswereatlogarithmicphaSe，themediumwasreplacedtothemediumdissolvedSJAMP．AccordingtothedensityofSJ姗bothHepG2ceilsandChangcellsweredividedintofourgroups：negativecontrolgroup(0lag／m1，groupA)，

4、SJAMPlow-dosegroup(0．51．tg／ml，groupB)，SJAMPmedium—dosegroup(2gtg／ml，groupC)andSJAMPhigh-dosegroup(8ug／ml，groupD)．AfterinterventionofSJAMP,observethecellsinelectronmicroscopy．MTTassaywasusedtoobservetheeffectofSJAMPonproliferationofHepG2andChangcells．Thece

5、llcyclewasmeasuredusingflowcytometry(FCM)．TheexpressionofPCNA，P53，MDM2，pRb，E2F一1，cyclinDl，CDK4andP21proteinwasdetectedbyimmunocytochemical。TheexpressionofP53，mdrll2，pRb，E2F一1，cyclinDlandCDK4mRNAwastestedbyreal-timePCR．Results：AfterinterventionofSJAMP,thei

6、nnerstructureofHepG2cellsWaschangedandapoptosisoccurred．MTTassayindicatedasignificantdose·aswell弱time·-dependentgrowthinhibitionwasobservedinSJAMP·-interventionHepG2cells(JPO．05)．Comparedwithnon—SJAM

7、Pinterventiongroup，moreceilsintheinterventiongroupsweretrappedatG1phase(P<0。05)andfewercaughtatSandG2phaSes(P0．05)．TheimmunocytochemicalresusltsshowedSJAMPdecreasedtheexpressionofPCNAproteinan

8、dincreaSedtheexpressionofp21proteinofHepG2cells，thedifferencesweresignificant(P<0．05)．Meanwhile，theexpressionofPCNAandP21proteinhadnosignificantdifferencesbetweeneachgroupofChangcells(P>O．05)．RT-PCRresultsshowedthat