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简介:Objective:Toinvestigatetheeffectsofanti-PML(promyelocyticleukemia)oranti-PML/RARa(promyelocyticleukemia/retionicacidreceptora)antisenseoligonucleotidesoncellgrowth,expressionofPML-RARamRNAandPML-RARa/PMLproteinlocationofNB4celllines.Methods:RT-PCRwasusedfordetectingPML-RARamRNAexpression,trypanblueexclusionforcellcount,methylcelloseassayforleukemiccolonyformingunitdetection,immuno-fluorescenceforPML-RARa/PMLproteinlocation.Results:Bothanti-PMLstartcodonregionantisence(STAS)andanti-PML-RARafusionregionantisence(FUAS)couldinhibitcellgrowthandtheformationofacutemyelocyticcolonyformingunitofcells(AML-CFU).Cellsbecomepartialdifferentiatedatdays5,beingmoreobviousinFUAS-treatedcellsthaninSTASones.DownregulationofPML-RARamRNAexpressionoccurredat24hoursinSTASandFUAS-treatedcellsandmaintainedforupto72hours.Immuno-fluorescenceanalysiswithanti-PMLmonoclonalantibodyshowedaremarkabledecreaseevencompletedisappearanceofmicrogranules.Theresidualgranulesbecameenlargedasdiscretedots(<10percell),similartonormalPODstructureinsomeSTAS-treatedcellsat24hours.At72hours,nearlyallthegranulesdisappeared.SimilarchangeswereobservedinFUAS-treatedcells.Conclusion:BothPMLandPML-RARaantisenceoligonucleotidescanspeciallyblocktheexpressionofPML-RARaatmRNAandproteinlevels.PMLproteinisimplicatedintheregulationsofcelldifferentiation.
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简介:Objective:Tostudythevaccinepotencyofgene-modifiedtumorcells.Methods:TheEL-4lymphomawastransducedwithrecombinantretroviruscontainingthemurineGM-CSFgeneorB7-1gene.Theeffectofgenetransductiononantitumorimmunitywasinvestigated.Results:Flowcytometryanalysisshowedthatexpressionoftheirsurfacemarkerbetweenwild-typeEL-4cellsandgenetransducedtumorcellswasthesameexceptforCD80positiveinB7-1genetransducedcells.GM-CSFgeneorB7-1genetransducedEL-4cellsresultedinremarkablelossoftumorigenicityinsyngeneticmice.ThesystemicprotectiveimmunitywasinducedagainstthechallengewithEL-4/wtcells.TherapeuticvaccinewithEL-4/GM-CSForEL/7-1cellscouldretardthegrowthofestablishedearly-stageEL-4/wttumorsignificantly,butnotretardthegrowthoflate-stageEL-4/wttumor.IrradiatedGM-CSFgenetransducedEL-4cellsshowedstrongvaccineeffectagainstEL-4cellchallenge,butirradiatedB7-1genetransducedEL-4cellsshowedweakvaccineeffect.RemarkablecooperativeantitumoreffectagainstEL-4cellchallengewasobservedwhenbothirradiatedEL-4/GM-CSFandEL-4/B7-1wereinoculatedtogether.Conclusion:GM-CSFgeneorB7-1genetransducedcombinationofthetwokindsofvaccinemayhavepotentialapplicationvalueinhumancancertreatment.
简介:目的:观察肝病患者PBMC中CD3+、CD4+、CD8+、CDl6+56+细胞亚群变化研究其在肝脏损伤中的意义。方法:流式细胞仪检测肝病患者PBMC中CD3+、CD4+、CD8+、CDl6+56广卜细胞亚群变化。结果:肝病患者除重型肝炎CD3+下降外,其他急慢性肝病组CD3+稍有上升。重型肝炎、慢乙肝活动期和急性肝炎CD4+下降而慢乙肝静止期、失代偿期肝硬化、恶性肿瘤CD4+下降不明显,而代偿期肝硬化CD4+反而上升。除肝硬化CD8+下降外,其他肝病组则上升。肝病CD4+/CD8+变化与CD4+相类似。除重型肝炎和慢乙肝CDl6+56+变化不明显,失代偿期肝硬化、急性肝炎、代偿期肝硬化和恶性肿瘤CDl6广卜56+下降。相关分析发现,肝病患者CDl6+56+与CD3+和CD4+/CD8+负相关,仅CD8+与急、慢性肝损伤指标有关。结论:PBMC中CD3+、CD4+、CD8+、CDl6+56+细胞亚群的检测在肝病的诊治应用意义有限,不宜仅单凭CD4+、CDl6+56+和CD4+/CD8+上升与否和CD8+下降与否来评判疾病和治疗效果,进一步结合其他亚群标志物可能更有利于疾病机制研制了解和疾病的诊断及治疗效果的评定。
简介:目的研究肝细胞癌(HCC)发生过程中细胞周期调控因子cyclinD1、CDK4和p16蛋白表达及其意义。方法应用免疫组织化学SP染色法检测正常肝组织、慢性肝炎,肝硬化,癌周肝硬化和肝癌组织中cyclinD1,CDK4和p16蛋白表达。对免疫组化结果进行计算机图象定量分析。结果cyclinD1,CDK4和p16蛋白的阳性单位(positiveunit,PU)和面数密度(areanumberdensity,NA)从慢性肝炎(PU分别为39.4、41.0和33.3;NA分别为236.7、272.7和237.4),肝硬化(PU分别为40.8、45.2和43.6;NA分别为313.8,354.6和322.9)癌周肝硬化(PU分别为55.5、59.4和54.4;NA分别为481.9、488.9和432.6)到肝癌(PU分别为59.6、63.7和58.1;NA分别为549.2、587.7和451.3)表达逐渐增强。癌周肝硬化和肝癌组织中cyclinD1、CDK4、p16的表达明显高于慢性肝炎和肝硬化(P值分别为0.034、0.020、0.030、0.007、0.003和0.005)但癌周肝硬化和肝癌组织之间差异无统计学意义,P值分别为0.433、0。535和0.447。慢性肝炎和肝硬化中cyclinD1、CDK4、p16阳性信号主要定位于胞核,而癌周肝硬化和HCC中主要定位于胞质。P16与HCC的分化程度有关,但未发现cyclinD1、CDK4与肿瘤分化程度之间有相关性。结论cyclin-细胞周期蛋白依赖性激酶(CDKs).细胞周期蛋白依赖性激酶抑制因子(CKIs)调控网络中,相关调控因子的异常可能参与了HCC的发生、发展。CyclinDl、CDK4的过度表达可能是肝癌发生过程中的早期事件。在HCC的发生过程中p16高表达可能是细胞周期正反馈调控的结果,在HCC的发生中可能属于早期事件,而p16表达下降或缺失可能是肝癌发生过程中的晚期事件。