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簡(jiǎn)介：MICROSATELLITEINSTABILITYAND/ORLOSSOFHETEROZYGOSITYINYOUNGGASTRICCANCERPATIENTSINITALYYIHHORNGSHIAO1,DANIELABOVO2,MARIAGUIDO2,CARLOCAPELLA3,MAUROCASSARO2,GRAZIELLABUSATTO2,VALENTINARUSSO2,4,ANGELOSIDONI5,ANNARPARENTI2ANDMASSIMORUGGE21LABORATORYOFCOMPARATIVECARCINOGENESIS,NCIFCRDC,NATIONALINSTITUTESOFHEALTH,FREDERICK,MD,USA2DEPARTMENTOFONCOLOGICALANDSURGICALSCIENCES,CATTEDRADIISTOCHIMICAEIMMUNOHISTOCHIMICAPATOLOGICA,IIICATTEDRADIANATOMIAPATOLOGICA,UNIVERSITA′DEGLISTUDI,PADOVA,ITALY3DEPARTMENTOFCLINICALANDBIOLOGICALSCIENCE,UNIVERSITYOFPAVIA,VARESE,ITALY4DEPARTMENTOFPATHOLOGY,UNIVERSITYOFCATANIA,CATANIA,ITALY5DEPARTMENTOFPATHOLOGY,UNIVERSITYOFPERUGIA,PERUGIA,ITALYGASTRICCANCERSARERARELYDIAGNOSEDBEFORETHEAGEOF40YEARSANDTHEINCIDENCEREACHESAPEAKDURINGTHE7THDECADEINTHEGENERALPOPULATIONAMOLECULARMECHANISMOFEARLYTUMORONSETMAYBEDETERMINEDBYCOMPARINGMICROSATELLITEINSTABILITYMSI,INDICATIVEOFERRORPRONEMISMATCHREPAIR,ANDLOSSOFHETEROZYGOSITYLOHBETWEENGASTRICCANCERSINPATIENTSI40YEARSOFAGEANDTHOSEOFOLDERAGESTHREETO5CHROMOSOMALLOCI,WHEREMSIAND/ORLOHARECOMMONLYFOUNDINGASTRICCANCERSINTHEGENERALPOPULATION,WEREEXAMINEDINFORMALINFIXED,PARAFFINEMBEDDEDSAMPLESFROM102PATIENTSI40YEARSOFAGEUSINGAPOLYMERASECHAINREACTIONBASEDNONRADIOACTIVESCREENINGMETHODMSIAND/ORLOHATAMINIMUMOF1LOCUSWEREDETECTEDIN11/102PATIENTSTHEFREQUENCYOFMSIAND/ORLOHATTHED11S904LOCUSWASSIGNIFICANTLYHIGHERTHANTHATATTHED2S119,D2S123,D5S409ANDIFNAREGIONSNOPREFERENTIALGENETICCHANGESATTHED11S904LOCUSWEREOBSERVEDINELDERLYPATIENTSAMONGSEVERALCLINICOPATHOLOGICALVARIABLES,ASTATISTICALLYSIGNIFICANTASSOCIATIONWITHMSIAND/ORLOHWASOBSERVEDONLYFORTUMORSLOCATEDATTHECARDIA,COMPAREDWITHTUMORSATTHEANTRUMANDTHECORPUSOURFINDINGSSUGGESTTHATAUNIQUEMECHANISMMAYBEINVOLVEDININCREASINGTHESUSCEPTIBILITYOFTHED11S904LOCUSFOREITHERMSIORLOH,ESPECIALLYFORCARDIATUMORSINYOUNGPATIENTSEARLYONSETOFGASTRICCANCERSINPATIENTSI40YEARSOFAGEISASSOCIATEDWITHGENETICCHANGESATPREFERENTIALCHROMOSOMALLOCI,INCLUDINGD11S904INTJCANCER8259–62,1999?1999WILEYLISS,INCALTHOUGHGASTRICCANCERINCIDENCEHASBEENDECLINING,ITWASSTILLRANKEDASTHE2NDMOSTCOMMONCANCERANDTHE2NDLEADINGCAUSEOFCANCERDEATHINTHEWORLDINTHE1990SPARKINETAL,1993BECAUSEOFTHEAGGRESSIVENATUREOFGASTRICCANCERS,THEOVERALL5YEARSURVIVALRATEISLESSTHAN20BREAUXETAL,1990GASTRICCANCERSUSUALLYOCCURINPATIENTSOLDERTHAN50YEARSOFAGEONLYABOUT5OFGASTRICCANCERPATIENTSAREYOUNGERTHAN40YEARSNEUGUTETAL,1996GASTRICCANCERSINPATIENTS?40YEARSOFAGEAREMOREAGGRESSIVETHANTHOSEINELDERLYPATIENTSFUJIMOTOETAL,1994COMPARISONOFGENETICALTERATIONSINGASTRICCANCERSBETWEENPATIENTS?40YEARSOFAGEANDOLDERPATIENTSMAYIDENTIFYASPECIFICMOLECULARMECHANISMASSOCIATEDWITHANEARLYONSETOFTUMORMICROSATELLITESARESHORTTANDEMLYREPEATEDDNASEQUENCESANDPRESENTTHROUGHOUTMAMMALIANGENOMESREPETITIVESEQUENCESOFDI,TRIANDTETRANUCLEOTIDEREPEATSAREPRESENTINMORETHAN50OFTHEHUMANGENOMEBECKMANANDWEBER,1992THETANDEMREPEATSOFDCDANAREPARTICULARLYABUNDANTALTERATIONSINTHENUMBEROFREPEATSPERSITE,KNOWNASMICROSATELLITEINSTABILITYMSI,HAVEBEENIMPLICATEDINVARIOUSHUMANDISEASES,INCLUDINGNEOPLASMSSPEICHER,1995MICROSATELLITESEQUENCESAREALSOHIGHLYPOLYMORPHICPOLYMORPHISMDUETODIFFERENTNUMBEROFREPEATSISVERYUSEFULTODETERMINEGENETICALTERATIONS,SUCHASLOSSOFHETEROZYGOSITYLOHINTHISSTUDY,WEEXAMINED5CHROMOSOMALLOCI,WHEREMSIAND/ORLOHARECOMMONLYFOUNDINGASTRICCANCERSINTHEGENERALPOPULATIONRHYUETAL,1994CHONGETAL,1994NAGELETAL,1995TAMURAETAL,1995LINETAL,1995SERUCAETAL,1995BUONSANTIETAL,1997,INGASTRICCANCERPATIENTS?40YEARSOFAGETHEFREQUENCYOFMSIAND/ORLOHANDTHEIRASSOCIATIONWITHOTHERCLINICOPATHOLOGICALDATAAREDISCUSSEDMATERIALANDMETHODSPATIENTSFORMALINFIXED,PARAFFINEMBEDDEDTISSUEBLOCKSFROM102PATIENTS?40YEARSOFAGEMEANAGE35YEARSRANGE16–40YEARSWERERETRIEVEDFROMTHEARCHIVESOFMANYITALIANHOSPITALSWITHCOMPARABLEGASTRICCANCERINCIDENCEDEMOGRAPHICANDPATHOLOGICALDATA,INCLUDINGAGE,GENDER,SITEOFTUMORANDTUMORSTAGEUSINGTHETNMSYSTEMBEAHRSANDMYERS,1983,WEREOBTAINEDFROMACOLLABORATIVEMULTICENTERSTUDYTHESITEOFTUMORWASCATEGORIZEDINTOTHEANTRUM,CORPUSORCARDIAOFSTOMACHNOCANCEREXTENDEDBEYONDTHEGASTRICESOPHAGEALJUNCTIONACCORDINGTOTHELAURE′NSYSTEM1965,GASTRICCANCERSWERECLASSIFIEDASEITHERINTESTINALORDIFFUSETYPEWHENBOTHPHENOTYPESCOEXISTED,CLASSIFICATIONOFTHETUMORTYPEWASBASEDONTHEMOSTREPRESENTATIVEHISTOLOGYGASTRITISNONATROPHICVSATROPHIC/METAPLASTICWASCLASSIFIEDUSINGTHEHOUSTONUPDATEDSYDNEYSYSTEMDIXONETAL,1996ALLCASESWEREJOINTLYASSESSEDBY2AUTHORSMCANDMRDNAEXTRACTIONNEOPLASTICANDADJACENTNONNEOPLASTICAREASWEREMICRODISSECTEDSEPARATELYFROMUNSTAINEDFORMALINFIXED,PARAFFINEMBEDDEDSECTIONSFORDEPARAFFINIZATION,PROTEINASEKDIGESTIONANDDNAPURIFICATION,ASDESCRIBEDPREVIOUSLYSHIAOETAL,1994MICRODISSECTEDNEOPLASTICAREASCONTAINEDATLEAST50TUMORCELLSWHENPOSSIBLE,LYMPHOCYTESWEREUSEDASANONNEOPLASTICCELLCONTROLPOLYMERASECHAINREACTIONPCRFIVECHROMOSOMALLOCIWITHCANDINUCLEOTIDEREPEATSD2S119,D2S123,D5S409,IFNAANDD11S904WERESELECTEDFORPCRPRIMERSD2S123AFM093XH3AANDAFM093XH3MD5S409AFM184YB6AANDAFM184YB6MIFNAIFNAPCR21ANDIFNAPCR22D11S904AFM081ZA5AANDAFM081ZA5MWEREOBTAINEDFROMTHEGENOMEDATABASEWEBSITEWWWGDBORGFORD2S119,UPPER5?CCAGTTTGGAA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