循环RNA检测与对胃癌诊疗的意义

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1、ABSTRACTobiectiveToestablishamethodofdetecting10wquaIltityofnucleicacidsbvnestedreal．timePCR，withwhichtodeten】1inethecirCulatingRNAinplaSmaorsupematantofcellculture．Todesignseveralculturemodels加1，所．D：serumdepriVation，hypoxia，lowtempera_tureandexposingwith锄ineoplasticagenttoanalyzethepot

2、entialfactorSthatenhancecirculatingRNAreleaSeincancerpatients，andmoreover，toinvestigatethesignific锄ceofdiagnosis锄dtherapyofgaStriccancer．MethodsCirculatingI矾AwaSisoI砷edwiththecombinationofTrizolrea鼯ntaIldcolumnmethod．Thenestedreal—timePCR，basedonSYBR呈∽enI，waLseStablished．HumangaStriccar

3、Cinomacellline(SGC7901)wascultllredunderserumdepriVation，hypoxiaandlowtemperatlJreconditions加’，f所DtoobservethechangeofsupematantI冰A，throu曲Whichtoanalyzethepossiblefactorsthata疏ctonthereleaSingofcirculatingRNAintumorpatients．SGCcellswerealsoeXposedt05．nuorouraciI(5．FU)andCisplatin(DDP)，t

4、heconcen仃ationofeXtracellularI之NAw嬲measuredat48h锄d96htoeValuatethechangeofcirCulatingRNAintumorpatientsanerthechemother印y加vf打．D．Inaddition，theexpressionofpactingeneincellularandex仃acellular鼢呛w嬲deteminedspecifically．Throu曲filtrationtestandexposuret0ribonuclease，theabili够ofanti-degradatio

5、nforcirculatingRNAw弱obseⅣed．PlasmaCXCR4andBmi一1mRNA，a11dCEAandCA19．9proteinswereeValuatedin89preoperatiVegaStriccancerpatientsand42healthysubjects．Si)(哆一nineofthecancerpatientswerealsoaSsessedaRersu唱e叮．Thecorrelationbe似eenthediagnosticUI江苏大学硕士学位论文ValuesofplaSmaCXCR4andBmi—lexpressionand

6、theleVeIsofCEAandCAl9—9waSinVestjgated．ResultsCirCulatingI矾Awassuccess如llyisolatedbythecombinationofTrizolreagentandcolumnmethod．Thesensitivityofnestedreal．timePCRislOo~1copies／山．Thequ锄tit)，ofeXtracellularRNAinthesupem雏mtofSGCcellswhichsumredserumdepriVing加dlowtemperatllredescendedcompa

7、ringwiththecontrolgroupwhileinthehypoxicculture，theex仃acellularRNAwasincreaSedsi印ificantlyaRer48h．Whenexposingtoanti-cancerdrugsfor48h，mosttllmorcellsdiedandaRerremoVingthedrugs，theextracellularRNAdecreasedsignmcantly．ThegeneexpressionofextraCellularRNAwassignificantlylowerthan