DOC

Expression

By Tony Riley,2014-07-18 02:27
10 views 0
Expression

    Expression

    Jan.2008,Volume5,No.1(SerialNo.38)JournalofUS.ChinaMedicalScience,ISSN1548?6648,USA

    ;ExpressionofKi67,PCNAinParotidTumors

    ;Q1UJia-xuan1.ZHUShengrongi.SudhottH2.HildmanH2

    ;f1.DepartmentofOralandMaxillofacialSurgery,TongjiHospital,TongjiMedicalCollegeofHuazhongUniversityofScience&

    ;Technology,Wuhan430030;2.DepartmentofENT-HeadandNeckSurgery,RuhrUniversity,Bochum4487&Germany)

    ;Abstract:ObjectiveToinvestigatetherelationbetweenthestainingofKi

    67andPCNAandtheclinical

    ;pathologicalgradeofparotidtumors.Methods32casesweredividedintothefollowingfourgroupsbasedon

    ;histologicaldiagnosis,theywerethenormalparotid,benignparotidtumor,Lowgradeandhighgradeparotid

    ;carcinoma.ThemethodofimmunohistochemicalmeasureswasusedtoinvestigateKi67,PCNAstainingin

    ;specimensofeachgroup,andthentheresultofeachgroupwascompared.ResultsInfourgroupstherateof

    ;positivestainingofKi

    67andPCNAinhighgradeparotidcarcinomawasthehighest,thelowgradeparotid

    ;carcinomawasthesecond.thebenignparotidtumorwasthethird,andthenormalparotidwasthelast.Themwas

    ;astatisticalsignificancebetweeneachtwogroupsinthepositivestainingofKi67andPCNAforthefourgroups

    ;rP<O.0l1.ConclusionThestainingofKi

    67andPCNAareusefulprognosticindicatorsforparotidtumors,and ;hasagoodcorrelationwiththeclinicalpathologicalgradeofparotidtumor.Themethodofimmunohistochemical

    ;stainingofKi67andPCNAcanbeusedinthediagnosisofparotidtumors. ;Keywords:parotidtumor;proliferatingcellnuclearantigen;Ki

    67;immunohistochemistry

    ;Ki

    67isacellproliferationassociatedantigenandanewkindofcellcycle.maintainingprotein.It’susedasa

    ;measureofproliferation….Proliferatingcellnuclearantigen(PCNA),isalsoappliedtolabelproliferatingcells.It

    ;wasshownthattheindexofKi

    67orPCNApositivecellscorrelatedwiththeirproliferatingpotential..Both ;Ki

    67andPCNAcouldbeappliedtothediagnosisandprognosisforthetumors,such

asthyroidcarcinoma,lung

    ;cancer,carcinomaofbladder,atypicalsmallglandularproliferationsoftheprostate.nonseminomatoustesticular

    ;germcelltumors.aciniccellcarcinomaoftheparotidglandandcarcinomaoftheextrahepaticbileduct..”.,but

    ;lessforparotidtumorinourknowledge.Wedetectedtheirexpressioninparotidtumorsinordertoinvestigatethe

    ;relationbetweentheexpressionofKi

    67.PCNAandpathologicalgradeofparotidtumors.

    ;MATERIALSANDMETHoDS

    ;1.TissueSpecimens

    ;32paraffinembeddedspecimenswere.

    ;obtainedfromDepartmentofENTHeadandNeckSurgery,Ruhr

    ;University,Germany,asidentifiedinadatabaseoutputfilethatincludedcasesfrom1997to2000.Theywere

    ;diagnosedbypathologistsandincludednormalparotid(n:8),pleomorphicadenoma(n:8),malignantmixed

    ;tumor(n:6),squamouscellcarcinoma(n:6),mucoepidermoidcarcinoma(n=2),andadenoidcysticcarcinoma

    ;(n:2).Tissuespecimenswerethendividedintothefollowingfourgroupsbasedonhistologicaldiagnosis:group

    ;A,normalparotid(n=8);groupB,pleomorphicadenoma(n:8);groupC,lowgr

adecarcinoma(n=8);groupD,

    ;ZHUShengrong(1952

    ),male,Ph.D.supervisor,directorofDepartmentofOralandMaxillofacialSurgery,Ton~iHospital

    ;TonalMedicalCollegeofHuazhongUniversityofScience&Technology;researchfield:oralmaxillofacialsurgery.

    ;37

    ;

    ;ExpressionofKi-67,PCNAinParotidTumo~

    ;highgradecarcinoma(n=8);patientagesrangedfrom22to89years(mean57.3years);male:19;female:13.

    ;2.ImmunohistochemicalStaining

    ;Thesampleswerefixedinl0%neutralbufferedformalinsolutionandembeddedinparaffinwax;serial

    ;sections(5m)werecutanddeparaffinatedinxyleneforantigenretrieval,sectionsweremicrowavedfor5minin

    ;citricbuffer(pH6.0)usingapressurecooker.Afterinactivationoftheendogenousperoxidaseby3%

    ;H2O2

    methanolfor10minandblockingofnonspecificantibodybinding(2-3drops

    ofgoatserum),thesections

    ;wereincubatedwiththeprimaryantiki67monoclonalantibody(x50,ZYM

EDHistostaintin-plusKitsLotNo:

    ;00350320California,USA)andwiththeprimaryantiPCNAmonoclonala

    ntibody(×1000,ZYMED

    ;Histostaintin-plusKitsLotNo:00350320California,USA)overnightat4?

    respectivelyinmoistchamber.After

    ;rinsingthreetimeswithTBSforatotalperiodof6min,thesectionswereincubatedwithbiotinylatedsecondary

    ;antibodyforl0mininmoistchamber.Followingthis,thesectionswererinsedthreetimeswithTBSforatotal

    ;periodof6min,andincubatedwithperoxidaseconjugatedstreptavidinfor

    10mininmoistchamber.Thesections

    ;wererinsedthreetimeswithTBSforatotalperiodof6min.Forlabelingcells,thesectionswereincubatedwith

    ;DABsubstratechromogensolutionfor10mininmoistchamber,thenrinsedwithTBSandaddedreagentD1,D2,

    ;D3(suppliedbyZYMEDHistostainTMPlusKits)tosections.Finally,thesectionswerecounterstainedusing

    ;Mayer’shematoxylin.Asanegativecontrol,theprimaryantibodywasreplacedwithnormalserum.

    ;3.AssessmentofImmunohistochemicalStaining

    ;Sectionswereassessedbyusingalightmicroscope(WestGermany).Thefieldsforcellcountingwere

    ;determinedusingarandomnumbertable.Aminimumof400tumorcells(groupA:fornormalparotidcells)were

    ;counted,allindexeswerecalculatedinpercent(%)asthenumberofcellsstainingpositive/totalnumberoftumor

    ;cellsrgroupA:fornormalcells).countedx100;thedistinctbrowngranularstainingofacell’snucleiwas

    ;consideredpositive.

    ;4.StatisticalAnalysis

    ;Theresultofpositivestainingisexpressedasthemean+SD.Kruskal-WallistestandstudentNewmanKeuls

    ;testareusedforstatisticalanalysis.

    ;RESUIrS

    ;TheresultsofpositiveimmunohistochemicalstainingofKi

    67andPCNAwereshowninTable1.The

    ;stainingofKi

    67wasgroupA>groupB>groupC>groupD.Thestatisticalsignificancewasshownbetweengroup

    ;AandB,BandC,CandD(P<0.01).ThestainingofPCNAwassimilarlyandshownthestatisticalsignificance

    ;betweengroupAandB,BandC,CandD(P<0.01).Undermicroscopethebrowngranularstainingwasinnuclei

    ;oftumorcellinKi

    67(Fig.1).Buttherewasalsobrowngranularstainingincytoplasmexceptinnucleioftumor

    ;cellmainlyinPCNA(Fig.21

    ;38

    ;Table1TheresultsofKi-67,PCNAimmunohistochemicalstaining(x?s,%) ;AntigenGroupAGroupBGroupCGroupD

    ;Ki670.73?0.336.47_+4-26l0.47?1.6822.3l+9.15

    ;PCNA0.80?0.59l0.06?5.40l8.50?3.9730.66_+9.8I

    ;

    ;ExpressionofKi-67,PCNAinParotidTumors

    ;Fig.1ImmunohistochemicalstainingforKi

    67inGroupA,B,C,D.a,b,c,disGroupA,B,C,D,

    ;respectively(originalmagnifications~200),A,B,C,D,isGroupA,B,C,D, ;respectively(originalmagnifications~400)

    ;39

    ;

    ;ExpressionofKi-67,PCNAinParotidTumors

    ;Fig.2lmmunohistochemicalstainingforPCNAinGroupA,B,C,D.a,b,C,disGroupA,B,C,

    ;D,respectively(originalmagnificationsx200),A,B,C,D,isGroupA,B,C, ;D,respectively(originalmagnificationsx400)

    ;

;ExpressionofKi-67,PCNAinParotidTumo~

    ;DISCUSSIoN

    ;Lossofheterozygosityatchromosome9p21,whichcontainsthetumor-suppressgenep16,acellcycle

    ;regulatorcouldresultinproliferativeantigenoverexpression131. ;Ki67,anapproximately300KDanuclearand

    ;nucleolarprotein,presentedinthenucleusduringtheS,G2

    MandlateG1,phasesofthecellcycle,isawidely

    ;usedmarkerofcellproliferation141.

    ;ThemonoclonalantibodyM1B1hasbeenreliablyusedtodetecttheKi67

    ;proteinI,15-171.

    ;PCNA,i.e.proliferatingcellnuclearantigen,isa36KDaacidicnuclearproteinthatisactivated

    ;duringtheG1phaseandearlySphaseofthecellcycle.PC10monoclonalantibodyagainstPCNAisavailableto

    ;labelproliferatingcellsinformalinfixed.paraffinembeddedhumantiss

    ueswithoutchemotoxicity.So,both

    ;Ki

    67andPCNAhavebeenusedinthediagnosisandprognosisformanykindsoftumors….Inourstudy,Ki67

    ;andPCNAimmunostainingwasused.Bothofthemwereshownthehighestpositivestaininginthehighgrade

    ;parotidcarcinomas.thesecondinthelowgradeparotidcarcinomas,theninthepleomorphicadenomas,thelastin

    ;thenormalparotidtissuesinthefourgroups,andthestatisticalsignificancewasshownbetweeneachtwogroups

    ;(P<0.01).So,Ki

    67andPCNAwaslinedtohistocytologicgradeofmalignantparotidtumors,andcouldbe

    ;appliedtodiscriminatingbetweenbenignandmalignanttumorsoftheparotidgland.Buttheimagesof

    ;immunohistochemicalstainingofcellsundermicroscopeinKi

    67weremoreclearlythannPCNA.We

    ;consideredthatthereasonwasthattheKi

    67immunohistochemistrywasobservedonlyinthenucleus,while ;PCNAwasdetectedinthemidG1phaseandSphase,thepositivereactionfor

    PCNAimmunohistochemistrywas

    ;observednotonlyinthenucleusbutalsointhecytoplasmtherefore,Ki

    67antigencouldbedetectedmore

    ;precisely.

    ;REFERENCES

    ;l

    Tsubochi,H.,Sato,N.,Hiyama,M.,eta1.Combinedanalysisofcyclooxygenase2expressionwithp53andKi67innonsmall

;celllungcancer.AnnThoracSurg,2006,82(4):I1981204.

    ;21

    ;3

    ;5

    ;l6

    ;7

    ;8

    ;9

    ;10

    ;Ishikawa,K.,Sakai,H.,Hosoi,M.,eta1.Evaluationofcellproliferationincaninetumorsbythebromodeoxyuridinelabeling

    ;method.immunostainingofKi

    67antigenandproliferatingcellnuclearantigen.JournalofToxicologicPathology,2006,,9(3):

    ;l23l27.

    ;Maddau,C..Confortini,M.,Bisanzi,S.,eta1.Prognosticsignificanceofp53andKi67antigenexpressioninsurgicallytreated

    ;nonsmallcelllungcancer:Immunocytochemicaldetectionwithimprint

    cytology.AmJClinPathol,2006,125(3):425-431.

    ;Kawahira,K.Immunohistochemicalstainingofproliferatingcellnuclearantigen(PCNA)inmalignantandnonmalignantskin

    ;diseasesArchDermatolRes,l999,291(78):4l34l8.

Report this document

For any questions or suggestions please email
cust-service@docsford.com