石蜡包埋组织中RNA提取方法的改进和完善

RNA550004RNA100RNATRIZOLRNA-20CRNARNARNART-PCRRNA!-actinRNA100RNA20~50mmoi/L260nm280nm1.75~2.0RT-PCRRNA190bp!-actinRNARNAmRNARNAR730.23A1000-2707200503-0241-03TheImprovementofIsolationTechnigueofRNAfromFormalin-fixedandParaffin-embeddedTissuesMENGOingYANGWen-xiuHUJunGAOOinDepartmentofPathologyGuiyangMedicalCollegeGuiyang550004ChinaAbstractObjectiveToimprovethetechnigueforextractingRNAfromformaiin-fixedandparaffin-embeddedtissue.MethodsSampiesof100casesthatdiagnosedpathoiogicaiiywerecoiiected.AftersomepretreatmentsuchasdeparaffinagewashingdigestiontotaiRNAwereextractedfromthesetis-sueswithTRIZOLreagentandthenkeptat-20Cforiongtimeabove2htosettieRNA.ThepurenessandcontentoftheseRNAextractswereassayedwithanucieicaciddetectinginstrumentandtheguaiityoftheextractswastestedwithdetectingofgene!-actinwithRT-PCR.ResultsThecon-centrationofRNAintheextractswerebetween20~50mmoi/L.Theratioof260nm/280nmwerebe-tween1.75~2.00.TheRNAof!-actinweredetectedinaiicases.ConclusionsThereasonabiepre-treatmentadoptedinthisstudymakesitfeasibietoextractRNAfromformaiin-fixedandparaffin-em-beddedtissueandtodetectgenesspecifictosomediseasesindaiiypathoiogicaidiagnosis.KeywordsRNAneopiasmparaffinembeddingreversetranscriptasepoiymerasechainreactionDNARNARNARNARNARNA11.119906~20031030202020101.2DEPCSigmaTRIZOLIn-vitrogenMMLVNEBMBIDNATAKARAdNTPKMBISPAN-14230320056JOURNALOFGUIYANGMEDICALCOLLEGEol30No32005.6ISHTrisSDSEDTA1.31.3.1RNA180C81tiptipPCR0.1%DEPC121121C22min1.3.2RNA175%0.1%DEPC0.1%DEPC5!m5~10RNA1.5mIRNAHE21.2mI8600r/min10min3EP37C15~20min8min31.2mI8600r/min10min3437C20min520mmOI/LTrispH8.020mmOI/LEDTApH8.02%SDSpH7.2200~250!IK400mg/L58C6K95C10min7TR-IZOL10min8200!I3min4C13000r/min15min9-20CRNA214C13000r/min15min101.2mI75%0.1%DEPC4C11000r/min10min118min120.1%DEPC58C10minRNARNA5~12RNA-85C1.3.3RNA1.3.3.14!IRNA0.5TBE100RNA260nm280nm1.3.3.2RT-PCR-actinRT-PCR190bpTGGAGAAATCTGGCACCACGAGGCGTACAGGGATAGCACRT-PCRMMLVPCR95C2min94C30s55C30s72C45s30RT-PCR2%0.5mg/L30min22.1100RNA8520~50mmOI/L260nm280nm1.751585mmOI/L260nm280nm1.45~1.6411RNA2.2RT-PCRRNA190bp-actin1MMarker1~23~45671RNAFig.1RNAisOIatedfrOmparaffin-embbedtissue31~3DNARNARNA4~6RNA24230RNARNARNATRIZOLRNARNAKl00mg/LRNA46RNAKRNAlRNARNARNAHERNARNARNAA260/A280l.75~2.00!-actinRNARNARNAmRNALMSRNARNA4lLiuHYeHDoganAetal.Tlll8g2lg2lisas-sociatedwithadvancedmucosa-associatedlymphoidtissuelymphomathatexpressesnuclearBcll0J.Blood200l984ll82-ll87.2HuangXZhangZLiuHetal.Tlll8g2lg2lingastricMALTlymphomaanddiffuselargeB-celllym-phomaofChinesepatientsJ.HematolJ200345342-345.3WillsTG.JadayelDMDuMOetal.Bcll0isinvolvedintll4p22g32ofMALTB-celllymphomaandmutatedinmutipletumortypesJ.Celll9999635-45.4TsujiSHisaokaMMonrimitsuYetal.DetectionofSYS-SSXfussiontranscriptioninsynovialsarcomabyreversetranscriptiontranscription-polymerasechainreactionusingarchivalparaffin-emdeddedtissueJ.AmJPazholl998l536l807-l8l2.5InagakiHOkabeMSetoMetal.API2-MALTlfusiontranscriptsinvolvedinmucosa-associatedlymphoidtissuelymphomamultiplexRT-PCRdetectionusingformalin-fixedparaffin-embeddedspecimensJ.AmJPathol200ll582699-706.6.SYT-SSXJ.200l306426-430.2004-09-2l2004-l2-202404lBaroneFcClarjRKPriceWJetal.Neuronspecificenolaseincreaseincerebralandsystemiccirculationfol-lowingfocalischemiaJ.BrainResl9936237.2.M..200l.l30-l423.Mg2+J.200320278-8l.2004-ll-l63423RNA石蜡包埋组织中RNA提取方法的改进和完善作者：孟青，杨文秀，胡军，高勤，MENGQing，YANGWen-xiu，HUJun，GaoQin作者单位：贵阳医学院,病理学教研室,贵州,贵阳,550004刊名：贵阳医学院学报英文刊名：JOURNALOFGUIYANGMEDICALCOLLEGE年，卷(期)：2005,30(3)被引用次数：2次参考文献(6条)1.魏永昆;孙孟红;朱红光滑膜肉瘤石蜡包埋组织SYT-SSX融合基因检测的临床病理意义[期刊论文]-中华病理杂志2001(06)2.InagakiH;OkabeM;SetoMAPI2-MALT1fusiontranscriptsinvolvedinmucosa-associatedlymphoidtissuelymphoma:multiplexRT-PCRdetectionusingformalin-fixedparaffin-embeddedspecimens2001(02)3.TsujiS;HisaokaM;MonrimitsuYDetectionofSYS-SSXfussiontranscriptioninsynovialsarcomabyreversetranscriptiontranscription-polymerasechainreactionusingarchivalparaffin-emdeddedtissue1998(06)4.WillsTG;JadayelDM;DuMQBcl10isinvolvedint(1;14)(p22;q32)ofMALTB-celllymphomaandmutatedinmutipletumortypes[外文期刊]19995.HuangX;ZhangZ;LiuHT(11;18)(q21;q21)ingastricMALTlymphomaanddiffuselargeB-celllymphomaofChinesepatients2003(05)6.LIUH;YeH;DoganAT(11,18)(q21,q21)isassociatedwithadvancedmucosa-associatedlymphoidtissuelymphomathatexpressesnuclearBcl102001(04)本文读者也读过(10条)1.齐妍.常彬.李洪安.王国亮.蒋金芳.梁伟华.李锋.QIYan.CHANGBin.LIHong-an.WANGGuo-liang.JIANGJin-fang.LIANGWei-hua.LIFeng一种简便的从石蜡包埋组织中提取RNA的方法[期刊论文]-生物技术通讯2007,18(1)2.孙冰.郭晓红.江泽飞.张峰.吴世凯.张利利.王敦梅.宋三泰.SUNBing.GUOXiao-hong.JIANGZe-fei.ZHANGFeng.WUShi-kai.ZHANGLi-li.WANGDun-mei.SONGSan-tai乳腺癌冰冻和石蜡包埋样本的RNA以及基因表达情况的比较研究[期刊论文]-临床肿瘤学杂志2010,15(7)3.马晓丽.黄淑红.汪运山.谢经武.张红卫病理组织中提取RNA的方法学研究[期刊论文]-山东大学学报（医学版）2004,42(5)4.赵志龙.宗志红.李建新.刘宏旭.赵惠儒.ZHAOZhilong.ZONGZhihong.LIJianxan.LIUHongxu.ZHAOHuiru从石蜡包埋肺癌组织提取RNA检测PML基因转录的研究[期刊论文]-中国肺癌杂志2008,11(4)5.曹慧仁.张元鑫.张红卫从石蜡包埋组织高效提取RNA的优化及Shh的RT-PCR检测[期刊论文]-山东大学学报(理学版)2004,39(4)6.程万宏.王艳俊.隗玉川石蜡包埋组织中提取RNA的方法改良[期刊论文]-中国中医药咨讯2010,02(7)7.陈柳青.曾学思.孙建方.ChenLiuqing.ZengXuesi.SunJianfang石蜡包埋皮肤组织标本中提取RNA进行逆转录PCR的研究[期刊论文]-中国麻风皮肤病杂志2006,22(1)8.吕杨.陈香美.尹忠.张雪光.王兆