核仁组成区嗜银蛋白AgNOR在子宫内膜病变组织中的定量研究

核仁组成区嗜银蛋白AgNOR在子宫内膜病变组织中的定量研究 核仁组成区嗜银蛋白AgNOR在子宫内膜病 变组织中的定量研究 第17卷第19期 2007年1O月 中国现代医学杂志ChinaJournalofModernMedicineVo1.17No.19 0ct.2OO7 文章编号:1005—8982(2007)19—2310—03 QuantitativestudyonAgNORinendometrialdiseases MAXiao—xin,JINYing—nan,ZHAOYan-hui,MIAOQing,ZHANGZhong—fu,LIXiao —han f1.DepartmentofObstetricsandGynecology,2.DepartmentofPathology,theSecondAffiliated HospitalofChinaMedicalUniversity,Shenyang,Liaoning110004,P.R.China) ? 论着? Abstract:【Objective】 ToinvestigatetheusageofAgNORinearlydiagnosisandprognosisofendometrialade— nocarcinoma.【Methods】 Experimentalgroup:107casesincluding25ofendometrialhyperplasia,19ofatypical proliferationand63ofendometrialadenocarcinoma.Controlgroup:25casesofnoYillalendometrium. Quantitative studyandmorphometricanalysisofAgNORsweretakenindifferentendometrialtissueswithsilver——stainingtech.. nique.【Results】 AgNORscounthadnostatisticaldifferences(P>0.05)innormalendometrium(2.51? 1.97),en— dometrialhyperplasia(2.92_+2.02)andatypicalproliferation(3.04_+2.o5).Inendometrialadenocarcinoma,AgNORs countincreasedobviously(7.43_+2.18),whichwashigherthanbenigndiseases[endometrialhyperplasia(2.92_+2.02), atypicalproliferation(3.O4?2.o5)]andcontrolgroup(2.51? 1.97)prominently(P<0.o5).AgNORscountincreased withgradeofcellulardifferentiation(4.69_+2.32),(5.38_+2.99),(7.54_+2.09),clinicalphase(3.68_+3.16),(4.53_+2.94), (7.27?2.84),(7.84_+2.33)andlymphaticmetastasis(7.68?2.42),(3.62? 3.27)(P<0.05).AgNORsmorphometrics changedfromsimpletypetodispersedtypewiththeprogressionofendometrialdiseases.【Conclusions】Thequanti tafivestudyandmorphometricanalysisofAgNORswerecorrelatedtotheincidenceofendometrialadenocarcinoma, anditcouldprovideevidencefordiagnosisofendometrialadenocarcinomaandcouldbeusedasaprognosticindex inmalignancies. Keywords:endometfialdiseases;AgNOR;endometrialadenocarcinoma CLCNumber:R737.33Documentcode:A 核仁组成区嗜银蛋白AgNOR在子宫内膜 病变组织中的定量研究术 马晓欣,金英楠,赵艳辉,苗青,张忠福,李晓晗 (中国医科大学附属盛京医院1.妇产科,2.病理科,辽宁沈阳ll0004) 摘要:目的探讨核仁组成区嗜银蛋白(AgNOtL)在子宫内膜腺癌早期诊断与预后中 的作用.方法采 用银染法染色,检测子宫内膜增殖症25例,非典型增生内膜19例,子宫内膜腺癌63 例,共107例为观察组;因 患宫颈原位癌子宫切除后的正常子宫内膜组织25例为对照组;计数正常子宫内膜 及不同内膜病变组织中单位 细胞核内AgNOtL的定量值,并对其进行形态学观察.结果AgNO1L每核颗粒数在正常子宫内膜(2.51? 1.97),子宫内膜增殖症(2.92?2.02),非典型增生内膜(3.04?2.05)间相比无统计学意义(P>0.05);在子宫 内膜腺癌(7.43?2.18)中,AgNOR颗粒数明显升高,与良性病变组(子宫内膜增殖症,非典型增生内膜)及对 照组相比有统计学意义(P<0.05);AgNOtL颗粒计数与子宫内膜腺癌的分化程度『高分化(4.69?2.32),中分化 (5.38?2.99),低分化(7.54?2.09)1,临床分期[I期(3.68?3.16),II期(4.53?2.94),III期(7.27?2.84),IV期 (7.84?2.33)],淋巴结转移[无转移(3.62?3.27),有转移(7.68?2.42)]密切相关(P<0.05);AgNOtL颗粒的形态分布 随着子宫内膜病变的进展由单一型转变为弥散型.结论AgNOtL的每核颗粒数及其形态分布与子宫内膜腺 癌的发生,发展密切相关;检测AgNOtL的定量值对早期诊断子宫内膜腺癌,判断疾病预后有一定的临床价值. 关键词:AgNOtL;子宫内膜病变;子宫内膜腺癌 中图分类号:1L737.33'文献标识码:A Receiveddate:Oct.14,2006 基金项目:该课题获辽宁省2002年博士启动基金(20021036) ? 2310? 第19期马晓欣,等:核仁组成区嗜银蛋白AgNOR在子宫内膜病变组织中的定量研究 Cellularabnormalproliferation1san~mportant biologicalcharacteroftumor.Argyrophilicnucleolar organizerregion(AgNOR)iscorrelatedtothesynthe. sisofribosomeandprotein,andthenumberandmor- phologyofAgNORsrepresentproliferativecellularac. tivity,thusitcanbeusedfordetectingmalignancy andgradingofthetumor.Nowadays,therolesofAg. NORsarewell—establishedinthediagnosisofbreast cancer,cervicalcancerandovarianepithelialtumor. AgNORhasbeenstudiedinendometrialadenocarci- noma.butonlyafewstudiesareavailable.Thepur- poseofthisstudywastoevaluatetheusageofAg- NORproteincountsandmorphometricanalysisvisu. alizedbyAgNORtechnique,inconformingdiagnosis andpredictingclinicaloutcomeofpatientswithen. dometria1adenocarcinoma. 1Materialsandmethods .1Materialsandreagents Allsamplesweregainedbyhysteroscopy,diag. nosticcurettageandabdominalhysterectomyinthe SecondAmliatedHospitalofChinaMedicalUniversi- tyfrom1990to2003.Thecaseshadbeenexcluded f0rhepaticorrenaldiseasesanddidntreceivehor— monetherapyforatleast3months.Experimental group:25casesofendometrialhyperplasia,19cases ofatypicalproliferationand63casesofendometrial adenocarcinoma.Theagesofendometrialadenocarci- nomagrouprangedfrom34to76withmedianageof 62.AccordingtoFIGOstagingin1988,theendome. trialadenocarcinomacasesincluded31ofstage1,19 ofstage2.11ofstage3and2ofstage4.According tolymphaticmetastasis:16caseswereoflymphatic metastasisandtheremaining47werenot.According tocelldifferentiation.26caseswereofG1.18cases 0fG2and19casesofG3.Controlgroup:25casesof normalendometriumincludedproliferativeandsecre- toryendometrium,andthesecasesreceivedhysterec- tomvbecauseofcervicalcancerinsitu.A1ldiagnosis wasconfirmedbypathology.Allthespecimenswere fixedwith10%formaldehydeandembeddedinwax. Reagents:AgNORkitswereprovidedbyZhongshan BiologicTechniqueCo.LTD. 1.2AgNORstaining Serialsectionof4txm.cutfromblocksroutinely processedinparaffinWaX,AgNORstainingwasper. formedwithasolutionobtainedbydissolvinggelatine inaqueousformicacid1volume,mixedwithsilver nitratesolution2volumes.Sectionsweredewaxedin xyleneandethano1.postfixedfor3Ominin3:1abso- luteethanol—aceticacidsolutionandrehydrated.The slideswereleftinthedarkfor27,60minutesand rinsedindeionizedwaterthreetimes,andthenthey werewashedwith5%sodiumthiosulfatesolutionfor5 minutes.Inadditiontheslideswererinsedinethanol f100%1andxylene,andthesmearsweremountedwith acoverslip. .3Resultsdetermination Silver—.stainedslideswereexaminedunderalight microscope,withAgNORsasbrownorblackdotswith avell0wishbackgroundofnucleus.Thebrown/black dotswerecountedinthenucleolusandalsooutside thenucleolusinthenucleus.Thenumberofdotswas countedin100cellsandtheaveragewastakenfor eachcase. I.4Statisticalanalysis StatisticalanalysiswasperformedbySPSS(ver. sion12.0package1fort—test.P<0.05wasconsidered tohavestatisticdifference. 2Results 2.1AgNORscountinnormalendometriunand够 ferentendometrialdiseases,seetable1. Table1ThevaluesofAgNOIcountinnormalen— dometriumanddifferentendometrialdiseases ThevaluesofAgNORscounthadnostatistical differencesfP>O.051innormalendometrium(2.51? 1.97),endometrialhyperplasia(2.92~2.02)andatypi- calproliferationf3.04~2.05).Inendometrialadeno- carcinoma,AgNORscountincreasedobviously(7.43+ 2.18),whichwashigherthanbenigndiseases(endome. trialhyperplasia,atypicalproliferation)andcontrol groupprominentlyfP<0.05). ThevaluesoftheAgNORscountpresent:en' dometrialadenocarcinoma>atypicalproliferation> endometrialhyperplasia>normalendometrium. 2311? 中国现代医学杂志第17卷 2.2RelationshipbetweenAgNORscountanden. ~metrialadenocarcinoma,seetable2. Table2ValuesofAgNORscountindifferenthistopatho. 1ogicparametersofendometrialadenocarcinoma AgNORscountincreasedwithgradeofcellular differentiation(4.69_+2.32),(5.38_+2.99),(7.54_+2.O9), clinicalphase(3.68+3.16),(4.53-+2.94),(7.27-+2.84), (7.84_+2.33)andlymphaticmetastasisf7.68?2.42). (3.62+3.27)(P<O.05).Inaword,thevaluesofAg. NORScountinendometrialadenocarcinomawerec0r- relatedtoclinicalstage,pathologicdifferentiationand lymphaticmetastasis<0.05). 2.3DistributionandmorphometricchangeofAg- NORsinnormalendometriunanddifferenten dometrialdiseases AgNORsgranuleswerevisibleasbrownorblack dotsmainlylocatedinthenucleolusofendometrial cells.Theyweresimpletypeinnormalendometrium andbenignendometrialdiseases,usuallypresenting regularinsizeandshape,withclearborderline.e quablearrangement.Theyweredispersedtypeinma— lignantadenocarcinomaceils,presentinground,strip orirregularinshape,differentinsizeanddisorderin arrangement,somegranulesevenoverlappedbetween eachother. 3Discussion Humannucleolarorganizerregions(NORs)are locatedinthesecondconstrictionofacrocentricchro. mosomes(chromosome13,14,15,21,22),andthey areloopsofDNAwhichcontainribosomalRNAgenes01. NORsaretranscribedbyRNApolymeraseIandare 0ftalimportancefortheultimatesynthesisofDro rein【21.AgNORsareacidicproteinsassociatedtothe NORswhichareselectivelystainedbyasilverco11oid technique.TheNORswithactivitycouldbestained whileinactivityorlessactivitynot, thereforeAgNORs couldreflectthechromosomallocationofrRNAas wellasthestatusofcellproliferation. Inmalignanttumors,thecelInucleolusincreased depolymerization,polymerizationdysfunction , rDNA lncreasedtranscription,andthetumorcellsactively proliferation[31,allofwhichcouldincreasetheva1ues ofAgNORsinnucleus,presentingincreasedAgNOR granulesinasamenucleolus,withdifferentsizeand disorderedarrangementunderlightmicroscopy.The quantitativestudyofAgNORproteiniscorrelatedto therapidityofcellproliferationandthereisevidence ofrelationshipbetweenAgNORscountsandthe progressofmalignanttumors.InapplyingtheAgNOR techniquetotumorpathology,theAgNORscounting methodappearstobeimportantindiagnosisbetween benignandmalignanttumors.PolitiENstudiedtode. finethediagnosticvalueofAgNORsquantitativeana1. ysisintheevaluationofhepaticlesions【羽. GuskiH showedthediagnosticandprognosticvaluesofargy. rophilicnucleolarorganizerregions(AgNORs)in atypicalductalhyperplasia(ADH),ductalcarcinoma insitu(DCIs)andmicroinvasiveductalcarcinoma (MDCA)ofthebreast[~3.TerlikowskiSreportedacor- relationbetweenthenumberofAgNORgranulesand thedegreeofcervicalintraepithelialneoplasia(CIN)啕. ZergerogluSstudiedAgNORsinovarianepithelialtu. morsandfoundthattheAgNORscorecanbeusedas aprognosticindexinmalignanciesI7~. Inourpresentstudy,wehavetriedtoobservethe valuesofAgNORscountanditsmorphometricvaria. tionin132casesincludingnormalendometriumand differentendometrialdiseaseswithsilver—staining technique.TheresuhsshowedthatAgNORgranules mainlylocatedinthenucleolusofceils,thevaluesof AgNORscountineachcellincreasedwiththedeteri orationofendometrialdiseases.Inendometrialhyper plasiaandatypicalproliferation,thevaluesslightly increasedwith1"1ostatisticalsignificancecomparedbe. tweenthetwogroupsandwithcontrolgroup. Inen dometrialadenocarcinoma,itsvalueincreasedDromi. nently,whichwascorrelatedtothetypeofthetumor. thegradeofthetumor, andthelymphnodestatus withmetastasis.Whatismore,themorphometricsof (Continuedonpage2321) ? 2312? 第19期高冬玲,等:胶质母细胞瘤组织中TRAIL受体达的研究 DcR的表达调控是否是TRAIL选择性诱导凋 亡的一个重要机制还存在争议,LEBLANC认为这 主要是因为几乎所有的研究都是通过探测mRNA, 而不是通过观察细胞表面的表达蛋白所致.本实验 免疫组化染色显示DcR蛋白的表达在胶质母细胞 瘤和正常脑组织中差异存在显着性(P<O.O1),这说 明DcR有可能在TRAIL的选择性凋亡中起重要调 控作用.同时,本实验还采用RT—PCR方法对比了 胶质母细胞瘤组织中DcR在mRNA水平的表达.结 果发现DcR在转录水平普遍表达,表达量显着高于 蛋白质水平的表达(P<O.01).综合免疫组化结果, 不仅说明了DcR的表达在胶质母细胞瘤和正常脑 组织中存在差异,而且在胶质母细胞瘤中表达减少 的调控位于转录后水平.这说明对在转录水平普遍 表达的DcR,如果采取反义技术封闭其表达,有可能 成为胶质母细胞瘤凋亡治疗的又一个新的策略. 参考文献: f1]KAUFMANNSH,STEENSMADP.OntheTRAILofanew therapyforleukemia[J].Leukemia,2005,19(12):2195—2202. 【2JBOURALEXISS,FINDLAYDM,EVDOKIOUA.Deathtothe badguys:targetingcancerviaApo2L/TRAIL[J].Apoptosis,2005, 1O(1):35-51. 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(顾靓编辑) (Continuationofthearticleonpage2312) AgNORgranuleschangedfromsimpletypetodis- persedtypewiththeprogressionofdiseases.These phenomenarepresentedproliferativecellularactivity, andaffinitiverelatedtotheoccurrenceandprogres- sionofendometrialadenocarcinoma.Whatismore, AgNORtechniqueisverysimpleanddoesnotrequire specialpreservationorfixationoftissue,itcanbe performedonformalin-fixedparaffin-embeddedsee- tionwhichisroutineforthehistopathology.Therefore, theAgNORtechniquemaybecomeanimportantpa- rameterinordertostudytheaggressivenessofthetu- norandbeusedasanimportantindexfordetecting malignancyandgradingofendometrialadenocarcino- ma. References: [1]XULZ.TumorPathologyMethodology[M].Shan曲ai:PublishCom— panyofShanghaiMedicalUniversity,1997:532.Chinese [2]EGANMJ,CROCKERJ.Nucleolarorganizerregionsinpathology[J]. BrJCancer,1992,65(1):1-7. [3]UNDERWOODJCE,GIRlDD.Nucleolarorganizerregionsasdiag— nosticdiscriminantsformalignancy【JJ.JPathol,1988,155(2):95— 97. 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