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1、調(diào)控人腦膠質(zhì)瘤中ezrin表達(dá)的miRNAs篩選及鑒定徐小琳江海波盛莉莉葛瑞祥沈軍毛捷皖南醫(yī)學(xué)院第一附屬醫(yī)院弋磯山醫(yī)院循證醫(yī)學(xué)教研室皖南醫(yī)學(xué)院第一附屬醫(yī)院弋磯山醫(yī)院神經(jīng)外科摘要:目的:探討調(diào)控人腦膠質(zhì)瘤中ezrin的miRNAs篩選及鑒定。方法:針對(duì)ezrin的測(cè)序序列,在線使用TargetScan>PITA>MicroRNA.org>miRGatorv2.0^miRDB等5種生物信息學(xué)軟件預(yù)測(cè)分析可能調(diào)控ezrin的miRNA分子,運(yùn)用熒光素酶報(bào)告載體實(shí)驗(yàn)檢測(cè)篩選候選miRNA分子,RT-PCR檢測(cè)11例膠質(zhì)瘤
2、組織和10例正常組織中候選miRNA分子mRNA的表達(dá)。結(jié)果:5種生物信息學(xué)軟件預(yù)測(cè)出819個(gè)可能調(diào)控ezrin的miRNAs,根據(jù)評(píng)分結(jié)果同時(shí)結(jié)合目前國(guó)內(nèi)外研究成果篩選出miR-148a>-204、-205、-34b、-370、-376b、-377、-410、-495、-548a-3p>-630和-96等12個(gè)miRNAso雙熒光素酶質(zhì)粒與ezrin3'-UTR作用位點(diǎn)突變體實(shí)驗(yàn),證實(shí)hsa-miR-204與相應(yīng)作用位點(diǎn)突變體EZR204Mutant無(wú)明顯作用(P>0.05),RT-PCR結(jié)果顯示miR-204
3、在腦膠質(zhì)瘤組織中的表達(dá)低于正常腦組織,差異有統(tǒng)計(jì)學(xué)意義(P<0.01)。結(jié)論:膠質(zhì)瘤組織中miR-204可通過(guò)靶向調(diào)控ezrin表達(dá)發(fā)揮生物學(xué)作用。關(guān)鍵詞:miR-204;ezi'in;膠質(zhì)瘤;熒光素酶報(bào)告載體;牛物信息學(xué);作者簡(jiǎn)介:徐小琳(1973-),女,助理研究員,(電話)15056458756,(電子信箱)xxiaolin66@163.com;作者簡(jiǎn)介:盛莉莉,女,副主任醫(yī)師,(電子信箱)13605535185@163.com,通信作者;作者簡(jiǎn)介:毛捷,男,副主任醫(yī)師,副教授,(電子信箱)myw921@1
4、63.com,通信作者。收稿日期:2017-02-15基金:安徽省自然科學(xué)基金項(xiàng)目(1408085MH205;1708085MH231)ScreeningandidentificationofmicroRNAstargetingezrininhumangliomaXUXiaolinJIANGHaiboSHENGL訂iGERuixiangSHENJunMAOJieDepartmentofEvidence-basedMedicine,TheFirstAffiliatedHospitalofWannanMedicalC
5、ollege;Abstract:Objective:ToscreenandidentifymicroRNAstargetingezrininhumanglioma.Methods:On-linesoftware,includingTargetSean,PITA,microrna.org,miRGatorv2.0andmiRDB,wereusedtopredictpotentialmicroRNAscapableofrcgulatingezrinexpression.DualLuciferaseReporterAs
6、saySystemwasusedtodeterminetheluciferaseactivityofthecandidatemicroRNAgenes,andreal一timePCR,todetectthemRNAvaluesofthecandidatemicroRNAsinthesamplesobtainedfrom11casesofgliomaandbraintissuesfrom10heaIthysubjects.Results:ThecandidatemicroRNAs,includingmiR-148a
7、,-204,-205,-34b,-370,-376b,-377,-410,-495,-548a~3p,-630and-96wereselectedfrom819potentialmicroRNAsthatwerepredictedtoregulatetheexpressionofezrin.DNAconstructsexpressingthemiRNAprecursorswereco-transfectedwithpLUC-ezrin3’UTR,andtheresultsindicatedthatonlytran
8、sfectionwithmiR-204expressionvectorsignifiedntlydown-regulatedtheluciferaseactivitycomparedtotransfectionwithanemptyvector(P>0.05),whereasreal-timePCRdemonstratedthatmiR-204expressionwass