简介:Colorectalcarcinoma(CRC)isacommoncauseofmorbidityandmortalityworldwide.TwopathogenicpathwaysareinvolvedinthedevelopmentofadenomatoCRC.ThefirstpathwayinvolvesAPC/β-catenincharacterizedbychromosomalinstabilityresultingintheaccumulationofmutations.ThesecondpathwayischaracterizedbylesionsinDNAmismatchrepairgenes.AberrantDNAmethylationinselectedgenepromotershasemergedasanewepigeneticpathwayinCRCdevelopment.CRCscreeningisthemostefficientstrategytoreducedeath.SpecificDNAmethylationeventsoccurinmultistepcarcinogenesis.EpigeneticgenesilencingisacausativefactorofCRCdevelopment.DNAmethylationshavebeenextensivelyexaminedinstoolfromCRCandprecursorlesions.ManymethylatedgeneshavebeendescribedinCRCandadenoma,althoughnodefiniteDNAmethylationbiomarkerspanelhasbeenestablished.MultipleDNAmethylationbiomarkers,includingsecretedfrizzled-relatedprotein2,secretedfrizzled-relatedprotein1,tissuefactorpathwayinhibitor2,vimentin,andmethylguanineDNAmethyltransferase,havebeenfurtherinvestigated,andobservationshaverevealedthatDNAmethylationbiomarkersexhibitwithhighsensitivityandspecificity.ThesemarkersmayalsobeusedtodiagnoseCRCandadenomainearlystages.Realtimepolymerasechainreaction(qPCR)issensitive,scalable,specific,reliable,timesaving,andcosteffective.Stoolexfoliatedmarkersprovideadvantages,includingsensitivityandspecificity.AstoolqPCRmethylationtestmayalsobeanenhancedtoolforCRCandadenomascreening.
简介:Someantitumoractivitiesofcomponent(E),extractedfromtherootofFagopynumCymosum(Trev)Meisn(FCTM),haverecentlybeendiscoveredinvivoandinvitro.ThecomponentE(CE)’spatternofactionwithtumorcellularDNAatthemolecularpharmacologicallevelwasinvestigatedbymacromolecularsynthesisexperiment(MSE)andhumanDNAinteractionsystemestablishedinourlaboratory.Theexperimentsdemonstratedthat,invitro,theagentcouldmarkedlyinhibittheincorporationof3H-TdRintothecellularDNA,andtheIC50inP388leukemiacellandinSGC-7901cellwas17.86μg/mland110.4μg/ml,respectively.Theagent,atmg/mllevel,couldproduceanintercalationreversionpatternwithDNAwithinashorttime(2hours).Butwhentheintervalwasprolongedforover4hours,theactionchangedtointercalationirreversiblepattern.Accordingtotheseobservations,theauthorsinferthatCEinteractswithDNAintwoways-directlyandindirectly.Theindirectaction,especiallyinlowconcentr
简介:Objective:TolookforthefurtherevidenceforHPVL1HPV16E6,HPV18E6andEBVascarcinogenicfactorsinlaryngealcarcinoma.Method:weexaminedrepresentativenumbersofspecimensfromlaryngealcancerwithhighlysensitivePCRtechniqueforthepresenceofHPVL1andhigh-risktypesHPV16E6,HPV18E6andEBVLMP1.Results:UsingPCRdetection,7.3%sampleswereHPVL1positive,52.03%wereHPV16E6positive,30.89%wereHPV18E6positiveand9.13%wereEBVLMP1positive.ThelowincidenceofHPVL1andhighincidenceofHPV-16E6andHPV18E6genessuggestthatHPVmightbeintegratedintotumorcells.OurresultssupportaroleofHPV-16andHPV-18infectioninthepathogenesisoflaryngealcarcinomainChina.Conclusion:IntegrationofE6intohostgenomeandstableexpressionofthesegenesmaybeassociatedwiththecarcinogenesisoflaryngealcarcinoma.HPV-16andHPV-18maysynergisticallyfunctiononthepathogenesisoflaryngealcarcinoma.Ourresultssuggestanassociationoflaryngealcarcinogenesisandinfectionwiththehigh-riskHPVtypes16,HPV18andEBV.
简介:Cancercellsdifferfromnormalcellsinvariousparameters,andthesedifferencesarecausedbygenomicmutationsandconsequentialalteredgeneexpression.Thegeneticandfunctionalheterogeneityoftumorcellsisamajorchallengeincancerresearch,detection,andeffectivetreatment.Assuch,theuseofdiagnosticmethodsisimportanttorevealthisheterogeneityatthesingle-celllevel.Dropletmicrofluidicdevicesareeffectivetoolsthatprovideexceptionalsensitivityforanalyzingsinglecellsandmolecules.Inthisreview,wehighlighttwonovelmethodsthatemploydropletmicrofluidicsforultrasensitivedetectionofnucleicacidsandproteinmarkersincancercells.Wealsodiscussthefuturepracticalapplicationsofthesemethods.
简介:TheDNAcontentandmorphometricfeaturesofhepatocellularcarcinoma(HCC)andlivercelldysplasia(LCD),includingnucleararea,nuclearperimeter,nuclearmaximumdiameterandnuclearcirclediameter,werequantitativelydeterminedbymeansofimageanalysistechnology.Theresultsshowedthatincomparisonwithnormalhepatocytes,LCDhadamarkedlyincreasedDNAcontentandnuclearmorphometricparameters,butthevalueswerelowerthanthoseforHCC.LCDshowedaslightincreaseinnuclearatypiarepresentedbythenuclearirregularindex,whichwasalsolessthanHCC.ThefindingsindicatethatLCDmaybeaprecaneerouslesionofHCC,tothecellsinanabnormalproliferativestate.
简介:Rat-1cellsweretransfectedwithDNAfromhumanesophagealcancer2K,4K,6K,7K.8K.Thetransformingfociwereobtainedandthetransformingcelllineswereestablished.Thecelllinescanformlargercolonyinsoftagar.Thosenudemiceinjectedsubcutaneouslywiththecellssufferedfromlargerfibroussarcoma.Thisindicatesthatthecelllineshavecarcinogenicity.TheexperimentalresultssuggestthathumanDNAsequenceandhumanHa-rasspecial616Kb(BamHI)bandarepresentintheDNAofthetransformingcells.Theover-expressionofrasgeneproductsP21werefoundinthetissuesofexophagealcancer,thetissuesadjacenttotumorandthetransformingcells.
简介:Objective:Theelevatedincidenceofobesityhasbeenparalleledwithhigherrisksofbreastcancer.Highadiposityincreasesleptinsecretionfromadiposetissue,whichinturnincreasescancercellproliferation.Theinterplaybetweenleptinandestrogenisoneofthemechanismsthroughwhichleptininfluencesbreastcarcinogenesis.Anunbalancedestrogenmetabolismincreasestheformationsofcatecholestrogenquinones,DNAadducts,andcancermutations.ThisstudyaimstoinvestigatetheeffectofleptinonsomeestrogenmetabolicenzymesandDNAadductioninbreastcancercells.Methods:Highperformanceliquidchromatography(HPLC)wasperformedtoanalyzetheDNAadducts4-OHE1[E2]-1-N3adenineand4-OHE1[E2]-1-N7guanine.Reportergeneassay,realtimereversetranscriptionpolymerasechainreaction(realtimeRT-PCR),andWesternblotwereusedtoassesstheexpressionofestrogenmetabolizinggenesandenzymes:CytochromeP-4501B1(CYP1B1),Nicotinamideadeninedinucleotidephosphate-quinoneoxidoreductase1(NQO1),andCatechol-O-methyltransferase(COMT).Results:LeptinsignificantlyincreasedtheDNAadducts4-OHE1[E2]-1-N3adenineand4-OHE1[E2]-1-N7guanine.Furthermore,leptinsignificantlyupregulatedCYP1B1promoteractivityandproteinexpression.TheluciferasepromoteractivitiesofNQO1andmRNAlevelsweresignificantlyreduced.Moreover,leptingreatlyreducedthereporteractivitiesoftheCOMT-P1andCOMT-P2promotersanddiminishedtheproteinexpressionofCOMT.Conclusions:LeptinincreasesDNAadductlevelsinbreastcancercellspartlybyaffectingkeygenesandenzymesinvolvedinestrogenmetabolism.Thus,increasedfocusshouldbedirectedtowardleptinanditseffectsontheestrogenmetabolicpathwayasaneffectiveapproachagainstbreastcancer.
简介:DuckhepatitisBvims(DHBV)DNAwasdetectedindifferenttumorousnodulesofduckswithhepaticmulticentriccancerorintrahepaticmetastasisbySouthernblottechnique.Among7duckswithhepatocellularcarcinomaofmultipletumornodules,thehybridizationpatternofIntegratedDHBVDNAIndifferenttumorousnoduleswasidenticalin3casesanddifferentin2cases.OnecaseshowedasimilarhybridizationpatternintwotumorousnodulesandotheronewasnegativetorDHBVDNA.IntegratedDHBVDNAwasalsoidentifiedinametastaticlungcancerofduckswithhepatocellularcarcinoma.Thehybridizationpatternofmetastasisoflungswasasthesomeasthatinprimaryhepatocellularcarcinoma.ThesamediscretehybridizationbandsInthedifferenttumorousnodulesindicatethatthesenodulesmightarisefromonetransformedcell.ThedifferenthybridizationpatternsInvarioustumorousnodulesshowthatthesetumorousnodulesmightarisefromvarioustransformedcells.Theresultssuggestthatthehyb
简介:USING3PRIMERPAIRSTODETECTHBVDNAINLIVERTISSUESFROMHEPATOCELLULARCARCINOMASWITHPCR TECHNIQUEChenMing陈明;LuLing吕凌;YaoJilu姚集鲁;Peng...
简介:目的:探讨胸腔积液中细胞DNA的含量分析对良恶性胸腔积液的鉴别诊断价值。方法:46例胸腔积液患者,分为恶性和良性两组,采用全自动细胞DNA定量分析系统测定胸腔积液细胞DNA的含量,同时检测癌胚抗原(CEA)。结果:对于恶性胸腔积液,全自动细胞DNA分析诊断的敏感性为58.3%,特异性为100.0%,准确性78.3%,而CEA分别为54.2%、81.8%和67.4%。如两项指标中任何一项阳性,诊断的敏感性、特异性和准确性分别为87.5%,81.8%和84.8%;两项指标同时阳性,特异性可达100.0%。结论:全自动细胞DNA定量分析对胸腔积液的鉴别有重要的临床参考价值,联合胸腔积液的CEA检测,可以提高诊断的有效性。
简介:分支链DNA信号放大技术(bDNA)是一种基于探针杂交信号放大的检测系统,无需对mRNA纯化和反转录即可对mRNA做精确的定量检测,有灵敏度高、线性范围宽、准确性好和结果稳定可靠等优点,具有良好的临床科研应用价值。本文就bDNA技术的原理及其在CMV、HBV、HCV和HIV等病毒定量检测、基因表达分析以及肿瘤基因分析等方面的应用作一介绍。
简介:目的探讨DNA修复基因MGMT和hMLT1在乳腺癌组织中的表达与病人年龄,肿瘤大小、淋巴结转移、TNM分期、癌组织分级、ER和PR受体及5年生存率的关系。方法应用免疫组织化学法,检测40例乳腺癌组织中MGMT和hMLT1蛋白表达。结果40例乳腺癌组织中MGMT表达阴性9例(22.5%),hMLT1表达阴性12例(30%)。对照组良性乳腺疾病组织均表达MGMT和hMLT1。癌组织同时不表达MGMT和hMLT1的乳腺癌病人有7例(17.5%),其5年生存率明显低于同时表达MGMT和hMLT126例乳腺癌病人(69.0%)(P<0.05)。MGMT和hMLT1在乳腺癌组织的表达与病人年龄、肿瘤大小、淋巴结转移,TNM分期、癌组织分级和激素受体状态等临床指标均无关(P均>0.05)。结论MGMT和hML腺疾病组织均有表达,在乳腺癌组织中有22.5%和30.0%不表达。二种DNA修复基因表达阴性的乳腺癌病人预后较差,提示联合检测MGMT和hMLT1两种DNA修复基因可作为判断乳癌预后的指标.
简介:目的探讨肝细胞癌(HCC)患者术前HBVDNA含量对切除术后肝功能恢复的影响。方法收集我科2007年1月至2007年6月行手术切除治疗的原发性肝癌患者共217例,应用实时荧光定量PCR方法检测血清中HBVDNA,按照HBVDNA含量分为两组:A组(HBVDNA≥1.0×105拷贝/ml)68例,B组(HBVDNA〈1.0×105拷贝/ml)149例。严密观察患者术后肝功能变化情况,采用SPSS13.0软件进行统计分析。结果术后两组患者的肝功能变化存在着显著差异,A组的肝功能损伤程度高于B组。术后第1、3、7天,丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)数值A组明显高于B组,而前白蛋白数值则B组高于A组,两组差异有统计学意义(P〈0.05);术后第3、7天,总胆红素(TBIL)数值A组明显高于B组,两组差异有统计学意义(P〈0.05)。除术前凝血酶原时间(PT)、前白蛋白、TBIL、AST和术中肝门阻断时间外,术前HBVDNA浓度对术后第7天的血清TBIL水平有显著影响。结论HCC患者术前HBVDNA含量对于术后肝功能恢复有明显影响,对术前HBVDNA含量较高的患者应在治疗肝癌的同时给予抗病毒治疗。
简介:目的评价DNA分析在痰液涂片细胞学检查中作用和意义。方法对HE染色痰涂片褪色后行Feulgen染色,用HPIAS-1000型高清晰度彩色全自动分析系统作DNA分析。结果35例肺癌细胞及9例不典型支气管上皮细胞DNA均为异倍体(heteroploid,H)核型。两组细胞DNA二倍体,异倍体(2C/H)核型分别占37.1%和66.6%(P〈0.001),五倍体/非整倍体(5C/AN)核型分别占71.4%、0.0%(P〈0.001),非整倍体(AN)核型占97.1%、0.0%(P〈0.001);而增生期百分比(PF)分别占(83.8±19.7)%、(47.629±.5)%(P〈0.001)。癌细胞组29例非小细胞癌与6例小细胞癌的2C/H核型分别占44.8%和0.0%(P〈0.001),5C/AN占65.5%、100.0%(P〈0.001),非整倍体(AN)核型为96.5%、100.0%(P〉0.05);而PF值为(83.1±20.1)%、(95.0±2.0)%(P〈0.02)。结论痰液细胞DNA分析指标(倍体核型、增殖期百分比)可作为细胞学肺癌诊断的重要理论参考。