应用免疫组织化学方法评价组织芯片的可靠性

26420058()JournalofXipanJiaotongUniversity(MedicalSciences)Vol.26No.4Aug.20051,1,1,1,1,2,2(:1.,710004;2.,710061):82,,(ER)(PR),92.9%,,82ERPR61%58.5%;,,51.9%50.0%,,53.8%53.8%,,57.1%60.7%,(P0.75),1.5mm,,,,:;;;;:R73:A:167128259(2005)0420399204EvaluationofthereliabilityoftissuechipwithimmunohistochemistrybydetectingERandPRexpressioninhumanbreastcancerYangJun,SuBaoshan,WangKangmin,ZhaoShiping,QiangLei,WangYili,SiLsheng(1.DepartmentofPathology,SecondHospitalofXipanJiaotongUniversity,Xipan710004;2.InstituteofTumorResearch,CollegeofLifeScienceandTechnology,XipanJiaotongUniversity,Xipan710061,China)ABSTRACT:ObjectiveToexplorethereliabilityoftissuechip(tissuemicroarray)andthemethodstoimprovethereliability.MethodsAtotalof82formalin2fixed,paraffin2embeddedtissueblocksofbreastcancerspecimenswereretrieved,alongwiththeircorrespondinghematoxylinandeosin(H&E)2stainedslide,fromthearchivesoftheDepartmentofPathology,SecondHospitalofXi‘anJiaotongUniversity.Patenttissuemicoarrayerwasusedtopreparetissuechip.Immunohistochemistrywasperformedtoanalyzetheexpressionofestrogenreceptor(ER)andprogesteronereceptor(PR)onthetissuechip.ResultsMorethan92.9%tissuesamplesonthetissuechipwereanalyzed.Themorecoresampleswerepunchedfromeachoriginalblock,themorespecimenscouldbeanalyzed.ThepositiverateoftheexpressionofERandPRonconventionaltissuesectionwere61%and58.5%respectively.Whenonecoresamplewaspunchedfromeachoriginalblock,thepositiverateoftheexpressionofERandPRonthetissuechipwere51.9%and50.0%respectively.Whentwocoresampleswerepunchedfromeachoriginalblock,thepositiveratewere53.8%and53.8%respectively.Whenthreecoresampleswerepunchedfromeachoriginalblock,thepositiveratewere57.1%and60.7%respectively.However,therewasnostatisticdifferencebetweenthevaluesusingtheabovethreemethods(P0.75).ConclusionWiththesamepreparationstandardandqualityoftissuechips,when1.5mmcoretissuesampleswerepunchedfromtheoriginalblockstopreparetissuechips,therewerenostatisticdifferenceamongthepositiveratebyusingthethreemethodsinpunchingcoresamples,althoughthepositivecoincidenceratebetweentheexpressionofERandPRonconventionaltissuesectionandtissuechipwasontherisewiththeincreaseofthenumberofcoretissuespunchedfromoneoriginalblock.Itcouldbeconcluded:2004210228:2005203210:(No.2001K102G2)(No.YG200120):(19722),(),.:.()26thattissuechiptechniquecouldsubstituteforconventionaltissuesectiontechniqueinimmunohistochemistry,insituhybridization(ISH)andfluorescenceinsituhybridization(FISH),andonecoretissuepunchedfromeachoriginalblockcouldensurethereliabilityandhigh2throughputcharactersoftissuemicroarraytechnique.KEYWORDS:tissuechip(tissuemicroarray);breastcarcinoma;ER;PR;immunohistochemistry1998Kononen[1](tissuemi2croarray),(tissuechip),,,,,82(es2trogenreceptor,ER)(progesteronerecep2tor,PR),,,11.11.1.1822,10%(),,HE1.1.2HT21(,ZL00262393.5ZL00207528.8ZL02139474.1)()1.1.3ERPR()S2P1.21.2.1823:128,,1,28;226,,2,52;328,,3,84,82164;,24(),822,4()HT2167()(1,,41/):;82HE,,50%;,,1.5mm,23mm;41,;,5m,1g/L,,,81.2.2SP:,3%()H2O210min,(0.01mol/LpH6.0),5%()10min,,371h,,3710h,3710h,DAB,,.TBS,1.2.3,,10%ERPR,,,1;231.2.4,,2,P0.0522.1,,,(1),ERPR(23)0044,,,.92.9%,(1)167()Fig.1Tissuechip67(RowColumn)2ERFig.2ExpressionofERinmammarycancerspecimenPositivegranulesinnucleusofcarcinomaSP4003PRFig.3ExpressionofPRinmammarycancerspecimenPositivegranulesinnucleusofcarcinomaSP4001Table1EffectonthenumberofsampleswiththreetissuechippreparingmethodsGroupingSpecimen(sample)numberMarkerAnalyzablespecimen(sample)numberAnalyzablespecimen(sample)rate(%)128(28)ER27(27)96.4(96.4)PR26(26)92.9(92.9)226(52)ER26(49)100.0(94.2)PR26(50)100.0(96.2)328(84)ER28(80)100.0(95.2)PR28(79)100.0(94.0)2.2(2,3)82ERPR61%58.5%;,,51.9%50.0%,88.9%92.3%;,53.8%53.8%,92.3%96.2%;,57.1%60.7%,96.4%100.0%(P0.75)2ERTable2ComparisonofpositiverateofERinmammarycancerwithtraditionalmethodandtissuechipGroupingSamplenumberPositivenumberNegativenumberTotalnumberPositiverate(%)Coincidencerate(%)114132751.988.9(24/27)214122653.892.3(24/26)316122857.196.4(27/28)450328261.0100.0(82/82)Total946916357.796.3(157/163)2=0.90035,0.900P0.7503PRTable3ComparisonofpositiverateofPRinmammarycancerwithtraditionalmethodandtissuechipGroupingSamplenumberPositivenumberNegativenumberTotalnumberPositiverate(%)Coincidencerate(%)113132650.092.3(24/26)214122653.896.2(25/26)317112860.7100.0(28/28)448348258.5100.0(82/82)Total927016257.098.1(159/162)2=0.85797,0.900P0.750104()263,,,,Kononen[1],0.6mm,,0.6mm6001300,Hoos[2],,Ki267,p53pRB59Mucci[3]50CGASYN,Garcia[4],,[5],,2.0mm[6],[7],[8],,,,82ERPR,,,82ERPR(P0.75,23),,,ERPR,,ERPR88.9%92.3%92.3%96.2%96.4%100%[2,3],HT21,,92.6%,,HE,,,50%,,,,1.5mm,400020000,Kononen[1-3]0.6mm,,,,,,,,,,,1.5mm,,,,,[9],,,1.5mm,,,:[1]KononenJ,BubendorfL,KallioniemiA,etal.Tissuemicroar2raysforhighthroughputmolecularprofilingoftumorspecimen[J].Natmed,1998,4:844-847.[2]HoosA,UristMJ,StojadinovicA,etal.Validationoftissuemicroarraysforimmunohistochemicalprofilingofcancerspeci2mensusingtheexampleofhumanfibroblastictumors[J].AmJPathol,2001,158(4):1245-1251.[3]MucciNR,AkdasG,ManelyS,etal.Neur