gli2在膀胱癌中的表达和意义及对t24细胞vegf和mmp9表达的影响-expression and significance of gli 2 in bladder cancer and its influence on expression of vegf and m mp9 in t24 cells.docx

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2018-06-04 发布于上海
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gli2在膀胱癌中的表达和意义及对t24细胞vegf和mmp9表达的影响-expression and significance of gli 2 in bladder cancer and its influence on expression of vegf and m mp9 in t24 cells.docx

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gli2在膀胱癌中的表达和意义及对t24细胞vegf和mmp9表达的影响-expression and significance of gli 2 in bladder cancer and its influence on expression of vegf and m mp9 in t24 cells

关键词：膀胱癌；Gli-2基因；VEGF；MMP-9；细胞增殖Theexpression andsignificance ofGli-2inbladdercancercelllinesT24 anditsinfluencesonVEGFandMMP-9expressionAbstract Objective:ToinvestigatetherelationshipbetweenGli-2expressionand clinicopathologicalcharacteristicthroughdetectingtheexpressionofGli-2,a key geneofHedgehog signalingpathway,in bladdercancertissues.To observetheroleofGli-2oncellproliferationanditsunderlyingmechanisms inbladdercancercelllinesT24,anditsinfluenceontheproteinexpressions ofVEGFandMMP-9.Methods:(1)TheexpressionlevelsofGli-2in77casesofbladdercarcinomatissue specimensweredetectedbyimmunohistochemistry(IHC),andtheir clinicopathologicalsignificancewasanalyzed.(2)Gli-2expressionwas down-regulatedinbladdercancercelllinesT24bysmallinterferingRNA (siRNA).ThetotalRNAandproteinswereextractedrespectively.RT-PCR wasusedtodetectGli-2mRNAexpression,westernblottoanalyse expressionlevelsofGli-2,VEGFandMMP-9protein,CCK-8assayand flowcytometrytoevaluatecellproliferation,apoptosisandcellcycle, respectively.Results:(1)Gli-2expressionwasobservedin100%ofbladdercancertissues.Gli-2 expressionwasintimatelyassociatedwithpathologicalstage,andthe expression levels were significantly higher in high stage bladder cancercomparedwithlowstage(P＜0.001).Andtheexpressioninmuscleinvasivebladdercancerwassignificantlyhigherthanthatinnon-muscleinvasive bladdercancer(P0.001),however,onlymarginalsignificancewas observedbetweenGli-2expressionandtumorgrade(P=0.052).(2)After down-regulationofGli-2expressioninT24cellsbysiRNA,themRNA expressionlevelsofGli-2geneintheexperimentgroupweresignificantly decreased,andstatisticalsignificancewasobservedcomparedwiththe controlgroup(t=-3.031,P0.05).Inaddition,Gli-2-siRNAtreatmentin bladdercellsresultedinasignificantreductionofGli-2mRNAandprotein expression,andtheexpressionlevelsofVEGFandMMP-9proteinwere significantlyreduced.Gli-2expressionknockdownproducedthedecreased proliferationofbladdercancercellsthroughevaluatedCCK-8testresults(P0.001).Flowcytometrywasusedtodetectthe